Methods of treating metabolic disorders with fgf21 variants

ABSTRACT

Provided herein are methods of treating, preventing, and managing metabolic or cardiovascular disorders and methods of reducing cardiovascular risk with FGF21 protein variants, including Fc-FGF21 variant fusion proteins.

I. FIELD OF THE INVENTION

The present disclosure relates to methods of treating, preventing, andmanaging metabolic or cardiovascular disorders and to methods ofreducing cardiovascular risk with FGF21 protein variants, includingFc-FGF21 variant fusion proteins.

II. BACKGROUND OF THE INVENTION

Insulin resistance is common and plays a central role in thepathogenesis of multiple diseases. Central adiposity is an importantsource of inflammatory cytokines and non-esterified fatty acids, whichhave been demonstrated to have detrimental effects on insulinsensitivity. Metabolic disorders associated with insulin resistance andadiposity manifest in various forms such as diabetes, obesity,dyslipidemia, coronary heart disease, and NAFLD/NASH. Fibroblast growthfactor 21 (FGF21) is thought to act on liver and adipose tissue toimprove insulin sensitivity, lower triglycerides, and reduce adiposity.Therefore, an FGF21 analog with clinically acceptable dosing regimen isexpected to become a frontline therapy for these metabolic disorders.

FGF21 is a member of the fibroblast growth factor family and hasmultiple metabolic functions. FGF21 is predominantly released fromhepatocytes and to a lesser extent from adipose and pancreatic β-cells.In mice, FGF21 has been shown to be a critical regulator of metabolismwhere expression is induced under conditions of both starvation andobesity. FGF21 induces various beneficial metabolic effects includingbody weight reduction, improved insulin sensitivity, increased highdensity lipoprotein-cholesterol (HDL-C), and decreased low densitylipoprotein-cholesterol (LDL-C), triglycerides (TG), and hepatic TGcontent in both nonclinical models and in patients with type 2 diabetesmellitus (T2DM). Furthermore, FGF21 effectively lowers glucose withoutinducing hypoglycemia in nonclinical animal models of diabetes. In T2DMpatients with increased body mass index, four week treatment with anFGF21 protein analog resulted in significant improvements in lipidprofiles, indicating the translatability of the metabolic effectsobserved in animal models to humans (Gaich et al., 2013, Cell Metab.18(3):333-40; Dong et al., 2015, Br J Clin Pharmacol. doi:10.1111/bcp.12676). Gaich et al. describes LY2405319, a variant ofFGF21, in a randomized, placebo-controlled, double-blindproof-of-concept trial in patients with obesity and type 2 diabetes,where patients received placebo or 3, 10, or 20 mg of LY2405319 dailyfor 28 days. Dong et al. describes a first-in-human study to evaluatethe pharmacokinetics/pharmacodynamics (PK/PD), safety and tolerabilityof single intravenous (IV) doses of PF-05231023, a long actingfibroblast growth factor 21 (FGF21) analogue, and report thatPF-05231023 levels peaked immediately post-IV dosing, with mean terminalhalf-lives of 6.5-7.7 h and 66.5-96.6 h for intact C- and N-termini,respectively.

There remains a need for improved FGF21 therapies with clinicallyacceptable dosing regimen useful for the treatment of metabolic diseasesand for reduction of cardiovascular risk in patients.

III. SUMMARY OF THE INVENTION

Novel modifications to FGF21 can improve the half-life and/or potencyover wild-type FGF21 to provide an FGF21 therapeutic for treating ormanaging (e.g., alleviating one or more symptoms of a disorder)metabolic or cardiovascular disorders or for reducing cardiovascularrisk, with clinically preferred dosing regimens.

In specific aspects, provided herein are FGF21 protein variants (forexample, FGF21 protein variants described in Table 1, e.g., such asFc-fusion proteins) and pharmaceutical compositions comprising suchFGF21 protein variants, for use in methods of treating, preventing,and/or managing hypertriglyceridemia and cardiac risk, insulinresistance such as patients with genetic mutations of insulin receptorand lipodystrophy, diabetes, obesity, and nonalcoholic fatty liverdisease (NAFLD)/nonalcoholic steatohepatitis (NASH).

V103 is a genetically engineered variant of human FGF21, stabilized viaintroduction of a novel disulfide bond, and fused at its N-terminus tohuman IgG1 Fc (see PCT Publication WO 2013/049247, which herebyincorporated by reference in its entirety). This combination ofstability and fusion of a human FGF21 mutant to IgG1 Fc results in anapproximately 50-100 fold improvement in half-life in cynomolgus monkeys(5 to 8 days) compared with wild-type (WT) human FGF21 (˜2 hours).

V103 has replicated published effects of FGF21 in animal models ofinsulin resistant T2DM including: reductions in body weight, glucose,insulin, serum TGs and liver TG content. V103 also demonstrated asignificant reduction in hepatic lipid, fibrosis and inflammation in amouse model of nonalcoholic steatohepatitis (NASH). Therefore, providedherein are pharmaceutical compositions comprising FGF21 protein variantV103 (SEQ ID NO: 11) for the treatment, prevention, and/or management ofmetabolic disorders including hypertriglyceridemia and cardiac risk,insulin resistance such as patients with genetic mutations of insulinreceptor and lipodystrophy, diabetes, obesity, and nonalcoholic fattyliver disease (NAFLD)/nonalcoholic steatohepatitis (NASH).

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing a metabolic disorder ora cardiovascular disorder in a human subject, wherein the human FGF21protein variant is provided for administration at a dose in the range of100 mg to 600 mg. In one aspect, provided herein is a human FGF21protein variant for use in a method of treating, preventing, or managinga metabolic disorder or a cardiovascular disorder in a human subject,wherein the human FGF21 protein variant is provided for administrationat an amount in the range of 100 mg to 600 mg.

In some embodiments of this aspect, the metabolic disorder orcardiovascular disorder is selected from hypercholesterolemia,dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease(NAFLD), nonalcoholic steatohepatitis (NASH), type 2 diabetes, andobesity.

In some embodiments of this aspect, treating, preventing, or managingthe metabolic disorder or cardiovascular disorder comprises or ischaracterized by reducing one or more of the following: body weight,liver fat content, elevated LDL-C, total-C, triglyceride, and Apo Blevels in the subject. In certain embodiments thereof, treating,preventing, or managing the metabolic disorder or cardiovasculardisorder comprises or is characterized by increasing HDL-C levels in thesubject. In some embodiments thereof, treating, preventing, or managingthe metabolic disorder or cardiovascular disorder comprises or ischaracterized by reducing triglyceride levels in the subject by at leastabout 40% or at least about 50%. In some embodiments thereof, treating,preventing, or managing the metabolic disorder or cardiovasculardisorder comprises or is characterized by reducing cardiovascular riskin the subject.

In some embodiments of this aspect, the human FGF21 variant is a fusionprotein comprising a human Fc region fused to a mature human FGF21protein or a fragment thereof comprising one or more mutations selectedfrom: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202Aaccording to the numbering of SEQ ID NO: 1. In certain embodimentsthereof, the human FGF21 protein variant comprises the amino acidsequence of SEQ ID NO: 11.

In some embodiments of this aspect, the human FGF21 protein variant isprovided for administration at a dose of at least 100 mg, 150 mg, 200mg, 250 mg, 300 mg, 350 mg, or 400 mg. In some embodiments thereof, thehuman FGF21 protein variant is provided for administration at a dose ofapproximately 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg,170 mg, 180 mg, or 190 mg, optionally wherein the human FGF21 proteinvariant is provided at a dose of approximately 100 mg or 150 mg. In someembodiments thereof, the human FGF21 protein variant is provided foradministration at a dose of approximately 200 mg, 210 mg, 220 mg, 230mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320mg, 330 mg, 340 mg, or 350 mg, optionally wherein the human FGF21protein variant is provided at a dose of approximately 200 mg, 250 mg,or 300 mg.

In some embodiments of this aspect, the human FGF21 protein variant isprovided in a form for subcutaneous administration. In some embodiments,the human FGF21 protein variant is provided for administration once amonth or once every 4 weeks, once every 3 weeks, once every 2 weeks, oronce every week. In some embodiments thereof, the human FGF21 proteinvariant is provided in a form for administration once a month or onceevery 4 weeks, once every 3 weeks, once every 2 weeks, or once everyweek. In some embodiments thereof, the human FGF21 protein variant isprovided in a form to be administered subcutaneously.

In one aspect, provided herein is a human FGF21 protein variant in themanufacture of a medicament for the treatment, prevention, or managementof a metabolic disorder or a cardiovascular disorder, wherein the unitdose of the human FGF21 protein variant is in the range of 100 mg to 600mg. In one aspect, provided herein is a human FGF21 protein variant inthe manufacture of a medicament for the treatment, prevention, ormanagement of a metabolic disorder or a cardiovascular disorder, whereinthe amount of the human FGF21 protein variant is in the range of 100 mgto 600 mg.

In some embodiments of this aspect, the metabolic disorder orcardiovascular disorder is selected from hypercholesterolemia,dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease(NAFLD), nonalcoholic steatohepatitis (NASH), type 2 diabetes, andobesity.

In some embodiments of this aspect, treating, preventing, or managingthe metabolic disorder or cardiovascular disorder comprises or ischaracterized by reducing one or more of the following: body weight,liver fat content, elevated LDL-C, total-C, triglyceride, and Apo Blevels. In some embodiments thereof, treating, preventing, or managingthe metabolic disorder or cardiovascular disorder comprises or ischaracterized by increasing HDL-C levels. In some embodiments thereof,triglyceride levels are reduced by at least about 40% or at least about50%. In some embodiments thereof, treating, preventing, or managing themetabolic disorder or cardiovascular disorder comprises or ischaracterized by reducing cardiovascular risk.

In some embodiments of this aspect, the human FGF21 variant is a fusionprotein comprising a human Fc region fused to a mature human FGF21protein or a fragment thereof comprising one or more mutations selectedfrom: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202Aaccording to the numbering of SEQ ID NO: 1. In some embodiments thereof,the human FGF21 protein variant comprises the amino acid sequence of SEQID NO: 11.

In some embodiments of this aspect, the human FGF21 protein variant isprovided for administration at a dose of at least 100 mg, 150 mg, 200mg, 250 mg, 300 mg, 350 mg, or 400 mg. In some embodiments, the humanFGF21 protein variant is provided for administration at a dose ofapproximately 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, or 400 mg.In some embodiments, the human FGF21 protein variant is provided foradministration at a dose of approximately 100 mg, 110 mg, 120 mg, 130mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, or 190 mg, optionallywherein the human FGF21 protein variant is provided for administrationat a dose of approximately 100 mg or 150 mg. In some embodiments, thehuman FGF21 protein variant is provided for administration at a dose ofapproximately 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg,270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350mg, optionally wherein the human FGF21 protein variant is provided foradministration at a dose of approximately 200 mg, 250 mg, or 300 mg.

In some embodiments, the human FGF21 protein variant is provided foradministration once a month or once every 4 weeks, once every 3 weeks,once every 2 weeks, or once every week. In some embodiments, the humanFGF21 protein variant is provided in a form for administration once amonth or once every 4 weeks, once every 3 weeks, once every 2 weeks, oronce every week. In some embodiments, the human FGF21 protein variant isformulated for subcutaneous administration. In some embodiments, thehuman FGF21 protein variant is provided in a form to be administeredsubcutaneously.

In one aspect, provided herein is a method of treating, preventing, ormanaging a metabolic disorder or a cardiovascular disorder in a humansubject, wherein the method comprises administering a human FGF21protein variant at a dose in the range of 100 mg to 600 mg to thesubject. In one aspect, provided herein is a method of treating,preventing, or managing a metabolic disorder or a cardiovasculardisorder in a human subject, wherein the method comprises administeringa human FGF21 protein variant at an amount in the range of 100 mg to 600mg to the subject.

In some embodiments, the metabolic disorder or cardiovascular disorderis selected from hypercholesterolemia, dyslipidemia,hypertriglyceridemia, nonalcoholic fatty liver disease (NAFLD),nonalcoholic steatohepatitis (NASH), type 2 diabetes, and obesity.

In some embodiments, treating, preventing, or managing the metabolicdisorder or cardiovascular disorder comprises or is characterized byreducing one or more of the following: body weight, liver fat content,elevated LDL-C, total-C, triglyceride, and Apo B levels in the subject.In certain embodiments, treating, preventing, or managing the metabolicdisorder or cardiovascular disorder comprises or is characterized byincreasing HDL-C levels in the subject. In some embodiments, treating,preventing, or managing the metabolic disorder or cardiovasculardisorder comprises or is characterized by reducing triglyceride levelsin the subject by at least about 40% or at least about 50%. In someembodiments, treating, preventing, or managing the metabolic disorder orcardiovascular disorder comprises or is characterized by reducingcardiovascular risk in the subject.

In some embodiments, the human FGF21 variant is a fusion proteincomprising a human Fc region fused to a mature human FGF21 protein or afragment thereof comprising one or more mutations selected from: Q55C,R105K, G148C, K150R, P158S, S195A, P199G, and G202A according to thenumbering of SEQ ID NO: 1. In certain embodiments, the human FGF21protein variant comprises the amino acid sequence of SEQ ID NO: 11.

In some embodiments, the method comprises administering the human FGF21protein variant at a dose of at least 100 mg, 150 mg, 200 mg, 250 mg,300 mg, 350 mg, or 400 mg to the subject. In some embodiments, themethod comprises administering the human FGF21 protein variant at a doseof approximately 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg,170 mg, 180 mg, or 190 mg to the subject. In some embodiments, themethod comprises administering the human FGF21 protein variant at a doseof approximately 100 mg or 150 mg to the subject. In some embodiments,the method comprises administering the human FGF21 protein variant at adose of approximately 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg,260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg,or 350 mg to the subject. In some embodiments, the method comprisesadministering the human FGF21 protein variant at a dose of approximately200 mg, 250 mg, or 300 mg to the subject.

In some embodiments, the human FGF21 protein variant is administeredonce a month or once every 4 weeks, once every 3 weeks, once every 2weeks, or once every week. In some embodiments, the human FGF21 proteinvariant is administered once a month or once every 4 weeks, once every 3weeks, once every 2 weeks, or once every week. In some embodiments, themethod comprises administering the human FGF21 protein variant in a formsuitable for subcutaneous administration. In some embodiments, themethod comprises subcutaneously administering the human FGF21 proteinvariant. In some embodiments, the human FGF21 protein variant isadministered subcutaneously.

Each of the foregoing aspects and embodiments, as well as other elementsdescribed herein, may be combined in any manner without limitation.

IV. DETAILED DESCRIPTION OF THE INVENTION

Provided herein are methods and pharmaceutical composition for use intreating, preventing, or managing (e.g., alleviating one or moresymptoms of a disorder) a metabolic disorder or a cardiovasculardisorder, comprising administering to a subject in need thereof a humanFGF21 protein variant (for example, an Fc-FGF21 mutant fusion proteinsuch as V103 (SEQ ID NO: 11)) at a dose or an amount in the range of 100mg to 600 mg. In specific aspects, a human FGF21 protein variant, suchas V103 (SEQ ID NO: 11) is administered at a dose in the range of 200 mgto 400 mg. In specific aspects, a human FGF21 protein variant, such asV103 (SEQ ID NO: 11) is administered at a dose of at least 200 mg, atleast 300 mg, or at least 400 mg. In specific aspects, a human FGF21protein variant, for example, an Fc-FGF21 mutant fusion protein such asV103 (SEQ ID NO: 11) is administered at a dose in the range of 100 mg to600 mg, for example, 250 mg to 350 mg, once every 4 weeks (or once everymonth), or once every 3 weeks or once every 2 weeks.

In one aspect, non-limiting examples of metabolic disorders orcardiovascular disorders to be treated, prevented or managed, by themethods provided herein include hypertriglyceridemia, diabetes, e.g.,type 2 diabetes mellitus, obesity, type 1 diabetes mellitus,pancreatitis, dyslipidemia, nonalcoholic fatty liver disease (NAFLD),nonalcoholic steatohepatitis (NASH), insulin resistance,hyperinsulinemia, glucose intolerance, hyperglycemia, metabolicsyndrome, hypertension, cardiovascular disease, acute myocardialinfarction, atherosclerosis, peripheral arterial disease, stroke, heartfailure, coronary heart disease, kidney disease, diabetic complications,neuropathy, disorders associated with severe inactivating mutations inthe insulin receptor, and/or gastroparesis.

Terminology

The term “native” or “wild-type,” in reference to FGF21, refers tobiologically active, naturally-occurring FGF21, including biologicallyactive, naturally-occurring FGF21 variants. An exemplary human FGF21wild-type sequence has NCBI reference sequence number NP 061986.1, andcan be found in such issued patents as, e.g., U.S. Pat. No. 6,716,626B1,(SEQ ID NO:1).

Met Asp Ser Asp Glu Thr Gly Phe Glu His Ser Gly Leu Trp Val Ser  1               5                  10                  15 Val Leu Ala Gly Leu Leu Leu Gly Ala Cys Gln Ala His Pro Ile Pro             20                  25                  30 Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val Arg Gln Arg Tyr         35                  40                  45 Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His Leu Glu Ile Arg     50                  55                  60 Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser Pro Glu Ser Leu 65                  70                  75                  80 Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln Ile Leu Gly Val                 85                  90                  95 Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly Ala Leu Tyr Gly             100                 105                 110 Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg Glu Leu Leu Leu         115                 120                 125 Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His Gly Leu Pro Leu     130                 135                 140 His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro Ala Pro Arg Gly 145                 150                 155                 160 Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro Ala Leu Pro Glu                 165                 170                 175 Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val Gly Ser Ser Asp             180                 185                 190 Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser Pro Ser Tyr Ala         195                 200                 205  Ser  209 The corresponding mRNA sequence coding for the full-length FGF21polypeptide (NCBI reference sequence number NM_019113.2) is shown below(SEQ ID NO:2):

  1 ctgtcagctg aggatccagc cgaaagagga gccaggcact caggccacct gagtctactc 61 acctggacaa ctggaatctg gcaccaattc taaaccactc agcttctccg agctcacacc121 ccggagatca cctgaggacc cgagccattg atggactcgg acgagaccgg gttcgagcac181 tcaggactgt gggtttctgt gctggctggt cttctgctgg gagcctgcca ggcacacccc241 atccctgact ccagtcctct cctgcaattc gggggccaag tccggcagcg gtacctctac301 acagatgatg cccagcagac agaagcccac ctggagatca gggaggatgg gacggtgggg361 ggcgctgctg accagagccc cgaaagtctc ctgcagctga aagccttgaa gccgggagtt421 attcaaatct tgggagtcaa gacatccagg ttcctgtgcc agcggccaga tggggccctg481 tatggatcgc tccactttga ccctgaggcc tgcagcttcc gggagctgct tcttgaggac541 ggatacaatg tttaccagtc cgaagcccac ggcctcccgc tgcacctgcc agggaacaag601 tccccacacc gggaccctgc accccgagga ccagctcgct tcctgccact accaggcctg661 ccccccgcac tcccggagcc acccggaatc ctggcccccc agccccccga tgtgggctcc721 tcggaccctc tgagcatggt gggaccttcc cagggccgaa gccccagcta cgcttcctga781 agccagaggc tgtttactat gacatctcct ctttatttat taggttattt atcttattta841 tttttttatt tttcttactt gagataataa agagttccag aggagaaaaa aaaaaaaaaa901 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa The mature FGF21 sequence lacks a leader sequence and may also includeother modifications of a polypeptide such as proteolytic processing ofthe amino terminus (with or without a leader sequence) and/or thecarboxyl terminus, cleavage of a smaller polypeptide from a largerprecursor, N-linked and/or O-linked glycosylation, and otherpost-translational modifications understood by those with skill in theart. A representative example of a mature FGF21 sequence has thefollowing sequence (SEQ ID NO:3, which represents amino acid positions29-209 of full length FGF21 protein sequence (NCBI reference sequencenumber NP 061986.1)):

His Pro Ile Pro Asp Ser Ser Pro Leu Leu Gln Phe Gly Gly Gln Val                  5                  10                  15 Arg Gln Arg Tyr Leu Tyr Thr Asp Asp Ala Gln Gln Thr Glu Ala His              20                  25                  30 Leu Glu Ile Arg Glu Asp Gly Thr Val Gly Gly Ala Ala Asp Gln Ser          35                  40                  45 Pro Glu Ser Leu Leu Gln Leu Lys Ala Leu Lys Pro Gly Val Ile Gln      50                  55                  60 Ile Leu Gly Val Lys Thr Ser Arg Phe Leu Cys Gln Arg Pro Asp Gly  65                  70                  75                  80 Ala Leu Tyr Gly Ser Leu His Phe Asp Pro Glu Ala Cys Ser Phe Arg                  85                  90                  95 Glu Leu Leu Leu Glu Asp Gly Tyr Asn Val Tyr Gln Ser Glu Ala His             100                 105                 110 Gly Leu Pro Leu His Leu Pro Gly Asn Lys Ser Pro His Arg Asp Pro         115                 120                 125 Ala Pro Arg Gly Pro Ala Arg Phe Leu Pro Leu Pro Gly Leu Pro Pro     130                 135                 140 Ala Leu Pro Glu Pro Pro Gly Ile Leu Ala Pro Gln Pro Pro Asp Val 145                 150                 155                 160 Gly Ser Ser Asp Pro Leu Ser Met Val Gly Pro Ser Gln Gly Arg Ser                 165                 170                 175 Pro Ser Tyr Ala Ser              180 The corresponding cDNA sequence coding for the mature FGF21 polypeptide(SEQ ID NO:3) is shown below (SEQ ID NO:4):

  1 caccccatcc ctgactccag tcctctcctg caattcgggg gccaagtccg gcagcggtac  61 ctctacacag atgatgccca gcagacagaa gcccacctgg agatcaggga ggatgggacg 121 gtggggggcg ctgctgacca gagccccgaa agtctcctgc agctgaaagc cttgaagccg 181 ggagttattc aaatcttggg agtcaagaca tccaggttcc tgtgccagcg gccagatggg 240 gccctgtatg gatcgctcca ctttgaccct gaggcctgca gcttccggga gctgcttctt 301 gaggacggat acaatgttta ccagtccgaa gcccacggcc tcccgctgca cctgccaggg 360 aacaagtccc cacaccggga ccctgcaccc cgaggaccag ctcgcttcct gccactacca 421 ggcctgcccc ccgcactccc ggagccaccc ggaatcctgg ccccccagcc ccccgatgtg 481 ggctcctcgg accctctgag catggtggga ccttcccagg gccgaagccc cagctacgct 541 tcctga 

The terms “FGF21 protein variant,” “human FGF21 variant,” “FGF21polypeptide or protein variant,” “FGF21 mutant,” or any like terms, aredefined as comprising human FGF21 in which a naturally occurring (i.e.,wild-type) FGF21 amino acid sequence has been modified, e.g., in whichat least one amino acid of the wild-type protein has been substituted byanother amino acid, and/or removed. Additionally, the variants mayinclude N- and/or C-terminal truncations relative to the wild-type FGF21protein, or internal amino acid deletions relative to wild-type humanFGF21 protein. Generally speaking, a variant possesses some modifiedproperty, structural or functional, of the wild-type protein. Forexample, the variant may have enhanced or improved physical stability inconcentrated solutions (e.g., less hydrophobic mediated aggregation),enhanced or improved plasma stability when incubated with blood plasmaor enhanced or improved bioactivity while maintaining a favorablebioactivity profile.

Acceptable amino acid substitutions and modifications which constitutedifferences between the FGF21 polypeptide and protein variants andmutants of the methods provided herein and wild-type FGF21 include, butare not limited to, one or more amino acid substitutions, includingsubstitutions with non-naturally occurring amino acid analogs, andtruncations. Thus, FGF21 protein variants include, but are not limitedto, site-directed FGF21 mutants, truncated FGF21 polypeptides,proteolysis-resistant FGF21 mutants, aggregation-reducing FGF21 mutants,FGF21 combination mutants, FGF21 conjugates (e.g., fatty acid-FGF21conjugate, PEG-FGF21 conjugate) and FGF21 fusion proteins (e.g., Fcdomain fusion protein, human serum albumin fusion protein), as describedherein.

The variant may possess increased compatibility with pharmaceuticalpreservatives (e.g., m-cresol, phenol, benzyl alcohol), thus enablingthe preparation of a preserved pharmaceutical formulation that maintainsthe physiochemical properties and biological activity of the proteinduring storage. Accordingly, variants with enhanced pharmaceuticalstability relative to wild-type FGF21 have improved physical stabilityin concentrated solutions under both physiological and preservedpharmaceutical formulation conditions, while maintaining biologicalpotency. As a set of non-limiting examples, variants provided herein maybe more resistant to proteolysis and enzymatic degradation; may haveimproved stability; and may be less likely to aggregate, than theirwild-type counterparts. As used herein, these terms are not mutuallyexclusive or limiting, it being entirely possible that a given varianthas one or more modified properties of the wild-type protein.

In specific aspects, FGF21 protein variants are biologically activevariants of the wild-type FGF21 protein that exhibits one or morebioactivity of wild-type FGF21. Bioactivity of wild-type FGF21 that hasbeen reported previously includes, but is not limited to, the following:(i) FGF21 has been shown to induce insulin-independent glucose uptake;(ii) FGF21 has also been shown to ameliorate hyperglycemia in a range ofdiabetic rodent models; (iii) transgenic mice over-expressing FGF21 werefound to be resistant to diet-induced metabolic abnormalities, anddemonstrated decreased body weight and fat mass, and enhancements ininsulin sensitivity (Badman, M. K. et al. (2007) Cell Metab 5, 426-37);(iv) administration of FGF21 to diabetic non-human primates caused adecline in fasting plasma glucose, triglycerides, insulin and glucagonlevels, and led to significant improvements in lipoprotein profilesincluding a nearly 80% increase in HDL cholesterol (Kharitonenkov, A. etal., 2007, Endocrinology 148, 774-81); (v) FGF21 as an importantendocrine hormone that helps to control adaptation to the fasting state(Badman et al., 2009, Endocrinology 150, 4931; Inagaki et al., 2007,Cell Metabolism 5, 415); and (vi) FGF21 can modulate downstream markerswhich include, without limitation, glucose or 2-deoxy-glucose uptake,pERK and other phosphorylated or acetylated proteins or NAD levels.

The term “native Fc” refers to molecule or sequence comprising thesequence of a non-antigen-binding fragment resulting from digestion ofwhole antibody or produced by other means, whether in monomeric ormultimeric form, and can contain the hinge region. The originalimmunoglobulin source of the native Fc is preferably of human origin andcan be any of the immunoglobulins, although IgG1 and IgG2 are preferred.Native Fc molecules are made up of monomeric polypeptides that can belinked into dimeric or multimeric forms by covalent (i.e., disulfidebonds) and non-covalent association. The number of intermoleculardisulfide bonds between monomeric subunits of native Fc molecules rangesfrom 1 to 4 depending on class (e.g., IgG, IgA, and IgE) or subclass(e.g., IgG1, IgG2, IgG3, IgA1, and IgGA2). One example of a native Fc isa disulfide-bonded dimer resulting from papain digestion of an IgG (seeEllison et al., 1982, Nucleic Acids Res. 10: 4071-9). The term “nativeFc” as used herein is generic to the monomeric, dimeric, and multimericforms.

The term “Fc variant” refers to a molecule or sequence that is modifiedfrom a native Fc but still comprises a binding site for the salvagereceptor, FcRn (neonatal Fc receptor). International Publication Nos. WO97/34631 and WO 96/32478 describe exemplary Fc variants, as well asinteraction with the salvage receptor, and are hereby incorporated byreference for this purpose. Thus, the term “Fc variant” can comprise amolecule or sequence that is humanized from a non-human native Fc.Furthermore, a native Fc comprises regions that can be removed becausethey provide structural features or biological activity that are notrequired for the fusion molecules of the fusion proteins of theinvention. Thus, the term “Fe variant” comprises a molecule or sequencethat lacks one or more native Fc sites or residues, or in which one ormore Fc sites or residues has be modified, that affect or are involvedin: (1) disulfide bond formation, (2) incompatibility with a selectedhost cell, (3) N-terminal heterogeneity upon expression in a selectedhost cell, (4) glycosylation, (5) interaction with complement, (6)binding to an Fc receptor other than a salvage receptor, and/or (7)antibody-dependent cellular cytotoxicity (ADCC). Fc variants aredescribed in further detail hereinafter.

The term “Fc domain” encompasses native Fc and Fc variants and sequencesas defined above. As with Fc variants and native Fc molecules, the term“Fc domain” includes molecules in monomeric or multimeric form, whetherdigested from whole antibody or produced by other means. In someembodiments of the present invention, an Fc domain can be fused to FGF21or a FGF21 mutant (including a truncated form of FGF21 or a FGF21mutant) via, for example, a covalent bond between the Fc domain and theFGF21 sequence. Such fusion proteins can form multimers via theassociation of the Fc domains and both these fusion proteins and theirmultimers are an aspect of the present invention.

The term “modified Fc fragment”, as used herein, shall mean an Fcfragment of an antibody comprising a modified sequence. The Fc fragmentis a portion of an antibody comprising the CH2, CH3 and part of thehinge region. The modified Fc fragment can be derived from, for example,IgG1, IgG2, IgG3, or IgG4. FcLALA is a modified Fc fragment with a LALAmutation (L234A, L235A), which triggers ADCC with lowered efficiency,and binds and activates human complement weakly. Hessell et al. 2007Nature 449:101-104. Additional modifications to the Fc fragment aredescribed in, for example, U.S. Pat. No. 7,217,798, which isincorporated by reference for this purpose.

The term “acute myocardial infarction” refers to myocardial necrosisresulting from interruption of the blood supply to a part of the heart,for example, due to acute obstruction of a coronary artery. Theresulting ischemia and oxygen shortage, if left untreated for asufficient period of time, can cause damage or death (infarction) of theheart muscle tissue (myocardium).

The term “atherosclerosis” is a vascular disease characterized byirregularly distributed lipid deposits in the intima of large andmedium-sized arteries, causing narrowing of arterial lumens andproceeding eventually to fibrosis and calcification. Lesions are usuallyfocal and progress slowly and intermittently. Limitation of blood flowaccounts for most clinical manifestations, which vary with thedistribution and severity of lesions.

The term “cardiovascular diseases” are diseases related to the heart orblood vessels.

The term “cardiovascular risk” refers to a combination of factorsassociated with a higher probability of cardiovascular event (e.g.,stroke or heart attack) occurring. Such factors may include, but are notlimited to, weight, BMI, cholesterol level, blood pressure, triglyceridelevels, diet, exercise routine, age, sex, family history, obesity,diabetes, and/or other metabolic factors. Guidelines for assessing andmanaging cardiovascular risk have been described. For example, theAmerican College of Cardiology (ACC) and the American Heart Association(AHA) have collaborated with the National Heart, Lung, and BloodInstitute (NHLBI) and stakeholder and professional organizations todevelop clinical practice guidelines for assessment of cardiovascularrisk, lifestyle modifications to reduce cardiovascular risk, managementof blood cholesterol in adults, and management of overweight and obesityin adults (Stone et al., 2014, Journal of the American College ofCardiology, 63 (25 Part B) 2889-2934; DOI: 10.1016/j.jacc.2013.11.002,which is hereby incorporated by reference for this purpose). The WorldHealth Organization also provides guidelines for assessingcardiovascular risk, for example, individuals with persistent raisedblood pressure≥160/100 mm Hg, blood cholesterol≥8 mmol/1, establishedischemic heart disease, or diabetes with renal disease.

The term “coronary heart disease”, also called coronary artery disease,is a narrowing of the small blood vessels that supply blood and oxygento the heart.

The terms “diabetes” and “diabetic” refer to a progressive disease ofcarbohydrate metabolism involving inadequate production or utilizationof insulin, frequently characterized by hyperglycemia and glycosuria.The terms “pre-diabetes” and “pre-diabetic” refer to a state wherein asubject does not have the characteristics, symptoms and the liketypically observed in diabetes, but does have characteristics, symptomsand the like that, if left untreated, may progress to diabetes. Thepresence of these conditions may be determined using, for example, thefasting plasma glucose (FPG) test, the oral glucose tolerance test(OGTT), or hemoglobin Alc (Hb1c) test. For the FPG test and OGTT, bothusually require a subject to fast for at least 8 hours prior toinitiating the test. In the FPG test, a subject's blood glucose ismeasured after the conclusion of the fasting; generally, the subjectfasts overnight and the blood glucose is measured in the morning beforethe subject eats. A healthy subject would generally have a FPGconcentration between about 90 and about 100 mg/dl, a subject with“pre-diabetes” would generally have a FPG concentration between about100 and about 125 mg/dl, and a subject with “diabetes” would generallyhave a FPG level above about 126 mg/dl. In the OGTT, a subject's bloodglucose is measured after fasting and again two hours after drinking aglucose-rich beverage. Two hours after consumption of the glucose-richbeverage, a healthy subject generally has a blood glucose concentrationbelow about 140 mg/dl, a pre-diabetic subject generally has a bloodglucose concentration about 140 to about 199 mg/dl, and a diabeticsubject generally has a blood glucose concentration about 200 mg/dl orabove. While the aforementioned glycemic values pertain to humansubjects, normoglycemia, moderate hyperglycemia and overt hyperglycemiaare scaled differently in murine subjects. A healthy murine subjectafter a four-hour fast would generally have a FPG concentration betweenabout 100 and about 150 mg/dl, a murine subject with “pre-diabetes”would generally have a FPG concentration between about 175 and about 250mg/dl and a murine subject with “diabetes” would generally have a FPGconcentration above about 250 mg/dl. For the Hb1c test, Hb1c levels<5.7% are generally considered to be in the normal range, Hb1c levels inthe range of 5.7-6.4% are generally considered to be pre-diabeteslevels, and Hb1c levels at 6.5% or above are generally considered to bediabetes levels (see, e.g., American Diabetes Association (ADA),Practice Guideline, Diabetes Care, 2018 supplement 1, which is herebyincorporated by reference for this purpose).

The term “dyslipidemia” is a disorder of lipoprotein metabolism,including lipoprotein overproduction or deficiency. Dyslipidemias may bemanifested by elevation of the total cholesterol, low-densitylipoprotein (LDL) cholesterol and triglyceride concentrations, and adecrease in high-density lipoprotein (HDL) cholesterol concentration inthe blood.

The term “glucose intolerance” or “Impaired fasting glucose” (IFG) or“Impaired Glucose Tolerance” (IGT) is a pre-diabetic state ofdysglycemia that is associated with increased risk of cardiovascularpathology. The pre-diabetic condition prevents a subject from movingglucose into cells efficiently and utilizing it as an efficient fuelsource, leading to elevated glucose levels in blood and some degree ofinsulin resistance.

The term “heart failure”, also called congestive heart failure, is acondition in which the heart can no longer pump enough blood to the restof the body.

The term “hyperglycemia,” also called high blood sugar, refers to acondition characterized by a higher than normal amount of glucose (atype of sugar) in the blood. Hyperglycemia can be diagnosed usingmethods known in the art, including measurement of fasting blood glucoselevels.

The term “hypertension” or high blood pressure refers to a conditionthat is a transitory or sustained elevation of systemic arterial bloodpressure to a level likely to induce cardiovascular damage or otheradverse consequences. Hypertension has been arbitrarily defined as asystolic blood pressure (SBP) above 140 mmHg or a diastolic bloodpressure (DBP) above 90 mmHg, but can also be defined based on publishedclinical guidance. For example, the following guidance for bloodpressure has been described: normal SBP<120 mmHg or DBP<80 mmHg;Elevated SBP in the range of 120-129 mmHg or DBP<80 mmHg; hypertension(HTN) stage 1 SBP in the range of 130-139 mmHg or DBP 80-89 mmHg; andHTN SBP≥140 or DBP≥90.

The term “hypertriglyceridemia” refers to high (hyper-) blood levels(-emia) of triglycerides. Elevated levels of triglycerides areassociated with atherosclerosis, even in the absence ofhypercholesterolemia (high cholesterol levels), and predispose tocardiovascular disease (e.g., contributes to increasing cardiovascularrisk). Generally, serum triglyceride levels in the range of 150 to 199mg per dL [1.70 to 2.25 mmol per L] are considered borderline-high serumtriglyceride levels; serum triglyceride levels in the range of 200 to499 mg per dL [2.26 to 5.64 mmol per L] are considered high serumtriglyceride levels; and serum triglyceride levels in the range of 500mg per dL [5.65 mmol per L] or higher are considered very hightriglyceride levels. In certain aspects, these general guidelines may beadjusted based on updated clinical guidelines for clinicians.

The term “hypoglycemia”, also called low blood sugar, occurs when yourblood glucose level drops too low to provide enough energy for yourbody's activities.

The term “hyperinsulinemia” is defined as a higher-than-normal level ofinsulin in the blood.

The term “insulin resistance” is defined as a state in which a normalamount of insulin produces a subnormal biologic response.

The term “kidney disease” or nephropathy is any disease of the kidney.Diabetic nephropathy refers to damage of the kidneys caused by diabetes;in severe cases it can lead to kidney failure a major cause of morbidityand mortality in people with type 1 or type 2 diabetes mellitus.

The term “metabolic syndrome” refers to a cluster ofconditions—increased blood pressure, high blood sugar, excess body fataround the waist, and abnormal cholesterol or triglyceride levels—thatoccur together, increasing risk of heart disease, stroke and diabetes.For example, excess body fat around the waist, in most men, isassociated with a 40-inch waist or greater; high blood sugar isassociated with a blood sugar/glucose level of at least 110 milligramsper deciliter (mg/dl) after fasting; high triglycerides is associatedwith levels at least 150 mg/dL in the bloodstream; low HDL is associatedwith levels less than 40 mg/dl; and increased blood pressure isassociated with levels 130/85 mmHg or higher.

The term “myocardial infarction” (MI), also called heart attack, isdefined as the irreversible death (necrosis) of heart muscle due toprolonged lack of oxygen supply (ischemia).

The term “NAFLD” or “nonalcoholic fatty liver disease” is defined as acondition in which excess fat is stored in the liver. This buildup offat is not caused by heavy alcohol use. When heavy alcohol use causesfat to build up in the liver, this condition is called alcoholic liverdisease. Typically, there are two types of NAFLD: simple fatty liver andnonalcoholic steatohepatitis (NASH).

The term “NASH” or “nonalcoholic steatohepatitis” is defined as a formof nonalcoholic fatty liver disease (NAFLD) in which a patient hashepatitis—inflammation of the liver—and liver cell damage, in additionto fat in the liver. Inflammation and liver cell damage can causefibrosis, or scarring, of the liver. In some cases NASH may lead tocirrhosis or liver cancer.

The term “simple fatty liver,” also called nonalcoholic fatty liver(NAFL), is a form of NAFLD in which a patient has fat in the liver butlittle or no inflammation or liver cell damage. Simple fatty livertypically does not progress to cause liver damage or complications.

The term “pancreatitis” is inflammation of the pancreas.

“Obesity” is defined as abnormal or excessive fat accumulation thatpresents a risk to health. A crude population measure of obesity is thebody mass index (BMI=Body weight (kg)/[Height (m)]²), a person's weight(in kilograms) divided by the square of his or her height (in meters). Aperson with a BMI of 30 or more is generally considered obese. Ingeneral, a person with a BMI equal to or more than 25 is consideredoverweight for example for Caucasians, but this guideline can beadjusted for ethnic differences. For example, with ethnic adjustment, aBMI≥27.5 can be considered obese for Asian individuals (WHO ExpertConsultation, 2004, Lancet, 363(9403):157-63). Asian individuals may be,for example, subjects of Asian descent or Asian ancestry.

The term “peripheral arterial disease” or “PAD” refers to a conditioncharacterized by a narrowing of the peripheral arteries to the legs,stomach, arms, and head (most commonly in the arteries of the legs). PADis similar to coronary artery disease (CAD) in that both PAD and CAD arecaused by atherosclerosis that narrows and blocks arteries in variouscritical regions of the body. The narrowing of arteries is typically dueto plaque that can, over time, harden and narrow the arteries limitingthe flow of oxygen-rich blood to organs and other parts of the body.

The term “stroke” is any acute clinical event, related to impairment ofcerebral circulation that lasts longer than 24 hours, while a shorterduration is considered a transient Ischemic Attack (TIA). A strokeinvolves irreversible brain damage, the type and severity of symptomsdepending on the location and extent of brain tissue whose circulationhas been compromised.

The term “type 1 diabetes mellitus” or “T1DM” is a conditioncharacterized by high blood glucose levels caused by lack of insulin.This can occur when the body's immune system attacks theinsulin-producing beta cells in the pancreas and destroys them. Thepancreas then produces little or no insulin.

The term “type 2 diabetes” or “type 2 diabetes mellitus” or “T2D” or“T2DM” refers to is a metabolic disease that causes sugar to collect inthe blood stream and is characterized by high blood glucose levelscaused by either a lack of insulin or the body's inability to useinsulin efficiently. Blood sugar levels are measured in milligrams perdeciliter (mg/dL) or mmol/L.

The term “management” or “manage(s)” or “managing” is understood as themanagement and care of a patient for the purpose of combating a disease,condition or disorder, which does not result in a cure, but alleviationof one or more symptoms of a disease, condition or disorder and/orreduction in hospital stay time.

The term “prevention” or “prevent(s)” or “preventing” refers toprophylactic administration to a healthy subject or to a subject at riskof developing a disease, condition or disorder (e.g., a subject who isconsidered to be prediabetic) to prevent the development of one or moreconditions mentioned herein. Moreover, the term “prevention” can alsoinclude prophylactic administration to patients being in a pre-stage ofthe conditions to be treated.

The term “treatment” or “treat(s)” or “treating” is understood as themanagement and care of a patient for the purpose of combating a disease,condition or disorder, for example, the reduction or amelioration of theprogression, severity, and/or duration of a disease, disorder orcondition. In specific aspects, treatment as used herein may include,but are not limited to, one or more of the following in the context ofdiseases, disorders or conditions which can be treated by administeringFGF21 or FGF21 variant proteins provided herein: (i) reduction intriglyceride levels (blood/serum triglyceride levels), for example,reduction to within a manageable range or normal range as determined bya clinician or clinical guidelines; (ii) reduction in body weight, forexample, reduction to within a manageable range or normal range asdetermined by a clinician or clinical guidelines or by at least about5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%,40%, 50%, 55%, 60%, or more; (iii) increase in insulin-independentglucose uptake; (iv) reduction in blood/serum glucose levels (e.g.,reduction to within a manageable range or normal range as determined bya clinician or clinical guidelines or by at least about 5%, 10%, 20%,30%, 40%, 45%, 50%, 55%, 60%, 70%, 80%, 90%, or 95% or more); (v)improvement in lipoprotein profiles; (vi) increase in HDL cholesterol(HDL-C) levels (e.g., increase by at least about 5%, 10%, 20%, 30%, 40%,50%, 60%, 70%, 80%, 90%, or more); (vii) reduction in LDL cholesterol(LDL-C) and/or total cholesterol (total-C), for example reduction towithin a manageable range or normal range as determined by a clinicianor clinical guidelines or by at least about 5%, 10%, 20%, 30%, 40%, or50% or more; (viii) reduction in liver fat content, for examplereduction to within a manageable range or normal range as determined bya clinician or clinical guidelines or by at least about 5%, 10%, 20%,30%, 40%, or 50% or more; and (ix) reduction in complication rates.

The term “therapeutically effective amount” or “effective amount” refersto an amount of a drug or a therapeutic agent that will elicit thedesired biological and/or medical response of a tissue, system or ananimal (including man) that is being sought by a researcher orclinician. In the context of FGF21 and FGF21 variant proteins providedherein, the desired biological and/or medical response may be one ormore of the following: (i) reduction in triglyceride levels (blood/serumtriglyceride levels), for example, by at least about 5%, 10%, 20%, 30%,40%, 45%, 50%, 55%, 60%, 70%, 80%, 90%, or 95% or more; (ii) reductionin body weight, for example, by at least about 5%, 6%, 7%, 8%, 9%, 10%,11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, ormore; (iii) increase in insulin-independent glucose uptake; (iv)reduction in blood/serum glucose levels (e.g., by at least about 5%,10%, 20%, 30%, 40%, 45%, 50%, 55%, 60%, 70%, 80%, 90%, or 95% or more);(v) improvement in lipoprotein profiles; (vi) increase in HDLcholesterol (HDL-C) levels (e.g., increase by at least about 5%, 10%,20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more); (vii) reduction in LDLcholesterol (LDL-C) and/or total cholesterol (total-C), for example byat least about 5%, 10%, 20%, 30%, 40%, or 50% or more; and (viii)reduction in liver fat content, for example by at least about 5%, 10%,20%, 30%, 40%, or 50% or more. In certain embodiments, triglyceridelevels may be reduced by at least about 40% or at least about 50%. Incertain embodiments, triglyceride levels may be reduced by at leastabout 40%. In certain embodiments, triglyceride levels may be reduced byat least about 50%. In some embodiments, liver fat content may bereduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more. Insome embodiments, liver fat content may be reduced by at least about 5%.In some embodiments, liver fat content may be reduced by at least about10%. In some embodiments, liver fat content may be reduced by at leastabout 20%. In some embodiments, liver fat content may be reduced by atleast about 30%. In some embodiments, liver fat content may be reducedby at least about 40%. In some embodiments, liver fat content may bereduced by at least about 50%. In some embodiments, the liver fatcontent may be reduced to within a manageable range or normal range asdetermined by a clinician or clinical guidelines.

As used herein, the singular forms “a,” “an” and “the” include pluralreferences unless the content clearly dictates otherwise. Thus, forexample, reference to “an antibody” includes a mixture of two or moresuch antibodies.

As used herein, the term “about” or “approximately” refers to +/−20%,more preferably, +/−10%, or still more preferably, +/−5% of a value.

The terms “polypeptide” and “protein”, are used interchangeably andrefer to a polymeric form of amino acids of any length, which caninclude coded and non-coded amino acids, naturally and non-naturallyoccurring amino acids, chemically or biochemically modified orderivatized amino acids, and polypeptides having modified peptidebackbones. The term includes fusion proteins, including, but not limitedto, fusion proteins with a heterologous amino acid sequence, fusionswith heterologous and homologous leader sequences, with or withoutN-terminal methionine residues; immunologically tagged proteins; and thelike.

The term “fragment” as used herein refers to a physically contiguousportion of the primary structure of a biomolecule. In the case ofproteins, a portion is defined by a contiguous portion of the amino acidsequence of that protein and refers to at least 3-5 amino acids, atleast 8-10 amino acids, at least 11-15 amino acids, at least 17-24 aminoacids, at least 25-30 amino acids, and at least 30-45 amino acids. Inthe case of oligonucleotides, a portion is defined by a contiguousportion of the nucleic acid sequence of that oligonucleotide and refersto at least 9-15 nucleotides, at least 18-30 nucleotides, at least 33-45nucleotides, at least 48-72 nucleotides, at least 75-90 nucleotides, andat least 90-130 nucleotides. In some embodiments, portions ofbiomolecules have a biological activity. In the context of FGF21polypeptide, FGF21 polypeptide fragments do not comprise the entireFGF21 polypeptide sequence set forth in SEQ ID NO:3.

The terms “individual”, “subject”, “host”, and “patient” are usedinterchangeably and refer to any subject for whom diagnosis, treatment,or therapy is desired, particularly humans. Other subjects may includecattle, dogs, cats, guinea pigs, rabbits, rats, mice, horses, and thelike. In some preferred embodiments the subject is a human.

As used herein, the term “sample” refers to biological material from apatient. The sample assayed by the present invention is not limited toany particular type. Samples include, as non-limiting examples, singlecells, multiple cells, tissues, tumors, biological fluids, biologicalmolecules, or supernatants or extracts of any of the foregoing. Examplesinclude tissue removed for biopsy, tissue removed during resection,blood, urine, lymph tissue, lymph fluid, cerebrospinal fluid, mucous,and stool samples. The sample used will vary based on the assay format,the detection method and the nature of the tumors, tissues, cells orextracts to be assayed. Methods for preparing samples are well known inthe art and can be readily adapted in order to obtain a sample that iscompatible with the method utilized.

The term “pharmaceutically acceptable carrier” refers to a carrier foradministration of a therapeutic agent, such as antibodies or apolypeptide, genes, and other therapeutic agents. The term refers to anypharmaceutical carrier that does not itself induce the production ofantibodies harmful to the individual receiving the composition, andwhich can be administered without undue toxicity. Suitable carriers canbe large, slowly metabolized macromolecules such as proteins,polysaccharides, polylactic acids, polyglycolic acids, polymeric aminoacids, amino acid copolymers, lipid aggregates, and/or inactive virusparticles. Such carriers are well known to those of ordinary skill inthe art. Pharmaceutically acceptable carriers in therapeuticcompositions can include liquids such as water, saline, glycerol, and/orethanol. Auxiliary substances, such as wetting or emulsifying agents, pHbuffering substances, and the like, can also be present in suchvehicles.

Therapeutic Methods

Provided herein are methods and pharmaceutical composition for use intreating, preventing, or managing (e.g., alleviating one or moresymptoms of) a metabolic disorder or a cardiovascular disorder,comprising administering to a subject in need thereof a human FGF21protein variant (for example, an Fc-FGF21 mutant fusion protein such asV103 (SEQ ID NO: 11)) at a dose in the range of 100 mg to 600 mg, forexample, once every day, once every 2, 3, 4, 5, or 6 days, once everyweek, once every 2 weeks, once every 3 weeks, or once every 4 weeks (oronce every month). In specific aspects, the human FGF21 protein variant(for example, an Fc-FGF21 mutant fusion protein such as V103 (SEQ ID NO:11)) is administered at a dose in the range of 200 mg to 400 mg. Inspecific aspects, the human FGF21 protein variant (for example, anFc-FGF21 mutant fusion protein such as V103 (SEQ ID NO: 11)) isadministered at a dose of approximately 200 mg, 250 mg, 300 mg, 350 mg,or 400 mg, for example, once every week, once every 2 weeks, once every3 weeks or once every 4 weeks (or once a month). In specific aspects,the human FGF21 protein variant (for example, an Fc-FGF21 mutant fusionprotein such as V103 (SEQ ID NO: 11)) is administered once every 4 weeks(or once a month). In certain aspects, the human FGF21 protein variant,such as V103 (SEQ ID NO: 11) is administered once every 3 weeks, onceevery 2 weeks, or once every week.

In a specific aspect, provided herein is a method of treating,preventing, or managing (e.g., alleviating one or more symptoms of) ametabolic disorder or a cardiovascular disorder (e.g., obesity orhypertriglyceridemia), comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc-FGF21 mutantfusion protein such as V103 (SEQ ID NO: 11)) at a dose in the range of100 mg to 600 mg once every week or once every two weeks. In a specificaspect, provided herein is a method of treating, preventing, or managing(e.g., alleviating one or more symptoms of) a metabolic disorder or acardiovascular disorder (e.g., obesity or hypertriglyceridemia),comprising administering to a subject in need thereof a human FGF21protein variant (for example, an Fc-FGF21 mutant fusion protein such asV103 (SEQ ID NO: 11)) at a dose in the range of 100 mg to 600 mg onceevery three weeks or once every four weeks (or once a month).

In a specific aspect, provided herein is a method of treating,preventing, or managing (e.g., alleviating one or more symptoms of) ametabolic disorder or a cardiovascular disorder (e.g., obesity orhypertriglyceridemia), comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc-FGF21 mutantfusion protein such as V103 (SEQ ID NO: 11)) at a dose in the range of200 mg to 400 mg once every week or once every two weeks. In a specificaspect, provided herein is a method of treating, preventing, or managing(e.g., alleviating one or more symptoms of) a metabolic disorder or acardiovascular disorder (e.g., obesity or hypertriglyceridemia),comprising administering to a subject in need thereof a human FGF21protein variant (for example, an Fc-FGF21 mutant fusion protein such asV103 (SEQ ID NO: 11)) at a dose in the range of 200 mg to 400 mg onceevery three weeks or once every four weeks (or once a month).

In a specific aspect, provided herein is a method of treating,preventing, or managing (e.g., alleviating one or more symptoms of) ametabolic disorder or a cardiovascular disorder (e.g., obesity orhypertriglyceridemia), comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc-FGF21 mutantfusion protein such as V103 (SEQ ID NO: 11)), at a dose in the range of100 mg to 300 mg once every week or once every two weeks. In a specificaspect, provided herein is a method of treating, preventing, or managing(e.g., alleviating one or more symptoms of) a metabolic disorder or acardiovascular disorder (e.g., obesity or hypertriglyceridemia),comprising administering to a subject in need thereof a human FGF21protein variant (for example, an Fc-FGF21 mutant fusion protein such asV103 (SEQ ID NO: 11)) at a dose in the range of 100 mg to 300 mg onceevery three weeks or once every four weeks (or once a month).

In a specific aspect, provided herein is a method of treating,preventing, or managing (e.g., alleviating one or more symptoms of) ametabolic disorder or a cardiovascular disorder (e.g., obesity orhypertriglyceridemia), comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc-FGF21 mutantfusion protein such as V103 (SEQ ID NO: 11)) at a dose of at least about100 mg, 150 mg, 200 mg, 250 mg, 300 mg, or 350 mg once every week oronce every two weeks. In a specific aspect, provided herein is a methodof treating, preventing, or managing (e.g., alleviating one or moresymptoms of) a metabolic disorder or a cardiovascular disorder (e.g.,obesity or hypertriglyceridemia), comprising administering to a subjectin need thereof a human FGF21 protein variant (for example, an Fc-FGF21mutant fusion protein such as V103 (SEQ ID NO: 11)) at a dose of atleast about 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, or 350 mg once everythree weeks or once every four weeks (or once a month).

In one aspect, non-limiting examples of metabolic disorders orcardiovascular disorders to be treated, prevented or managed, by themethods provided herein include obesity, hypertriglyceridemia,dyslipidemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholicsteatohepatitis (NASH), diabetes, e.g., type 2 diabetes mellitus (T2DM)and type 1 diabetes mellitus (T1DM), disorders associated with severeinactivating mutations in the insulin receptor, pancreatitis, insulinresistance, hyperinsulinemia, glucose intolerance, hyperglycemia,metabolic syndrome, hypertension, cardiovascular disease, acutemyocardial infarction, atherosclerosis, peripheral arterial disease,stroke, heart failure, coronary heart disease, kidney disease, diabeticcomplications, neuropathy, and gastroparesis.

In one aspect, non-limiting examples of metabolic disorders orcardiovascular disorders to be treated, prevented or managed, by themethods provided herein include obesity, hypertriglyceridemia andcardiac risk, insulin resistance such as patients with genetic mutationsof insulin receptor and lipodystrophy, diabetes (e.g., T1DM or T2DM),and nonalcoholic fatty liver disease (NAFLD)/nonalcoholicsteatohepatitis (NASH).

In one aspect, provided herein is a method of treating, preventing, ormanaging a metabolic disorder (e.g., obesity) or a cardiovasculardisorder, comprising administering, e.g., s.c., to a subject in needthereof a human FGF21 protein variant (for example, an Fc fusion proteinsuch as V103 (SEQ ID NO: 11)) at a dose in the range of 100 mg to 600mg, or more specifically at a dose in the range of 100 mg to 300 mg or200 mg to 400 mg, for example, once every day, once every 2, 3, 4, 5, or6 days, once every week, once every 2 weeks, once every 3 weeks, or onceevery 4 weeks (or once every month). In one aspect, provided herein is amethod of treating, preventing, or managing a metabolic disorder (e.g.,obesity) or a cardiovascular disorder, comprising administering, e.g.,s.c., to a subject in need thereof a human FGF21 protein variant (forexample, an Fc fusion protein such as V103 (SEQ ID NO: 11)) at a dose ofapproximately 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg,170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg,260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg,or 350 mg, for example, once every day, once every 2, 3, 4, 5, or 6days, once every week, once every 2 weeks, once every 3 weeks, or onceevery 4 weeks (or once every month). In a particular aspect, providedherein is a method of treating, preventing, or managing a metabolicdisorder (e.g., obesity) or a cardiovascular disorder, comprisingadministering, e.g., s.c., to a subject in need thereof a human FGF21protein variant (for example, an Fc fusion protein such as V103 (SEQ IDNO: 11)) at a dose of approximately 300 mg, subcutaneously, every 4weeks (or every month). In a particular aspect, provided herein is amethod of treating, preventing, or managing a metabolic disorder (e.g.,obesity) or a cardiovascular disorder, comprising administering, e.g.,s.c., to a subject in need thereof a human FGF21 protein variant (forexample, an Fc fusion protein, such as V103 (SEQ ID NO: 11)) at a doseof approximately 300 mg, subcutaneously, every 3 weeks. In a particularaspect, provided herein is a method of treating, preventing, or managinga metabolic disorder (e.g., obesity) or a cardiovascular disorder,comprising administering, e.g., s.c., to a subject in need thereof ahuman FGF21 protein variant (for example, an Fc fusion protein, such asV103 (SEQ ID NO: 11)) at a dose of approximately 300 mg, subcutaneously,every 2 weeks. In a specific aspect, provided herein is a method oftreating, preventing, or managing a metabolic disorder (e.g., obesity)or a cardiovascular disorder, comprising administering to a subject inneed thereof a human FGF21 protein variant (for example, an Fc fusionprotein, such as V103 (SEQ ID NO: 11)) at a dose in the range ofapproximately 200 mg to 250 mg, subcutaneously, every 3 weeks. Inspecific aspects, methods provided herein are for treating a humansubject with hypercholesterolemia (e.g., primary hypercholesterolemia)or dyslipidemia (e.g., mixed dyslipidemia). In particular aspects,methods provided herein are for treating a human subject withhypertriglyceridemia, in particular, severe hypertriglyceridemia. Incertain aspects, methods provided herein are for treating a humansubject with type 2 diabetes or a human subject who is obese. Inspecific aspects, methods provided herein are for treating a humansubject 18 to 55 years of age, who has (i) a BMI within the range of30-45 kg/m², inclusive (with ethnic adjustment of greater than or equalto 27.5 kg/m² BMI for Asian subjects), and (ii) triglyceride levels inthe range of 150 to 500 mg/dL (1.69-5.65 mmol/L), prior toadministration of the human FGF21 protein variant.

In particular aspects, provided herein is a method of treating,preventing, or managing (e.g., alleviating one or more symptoms of)hypercholesterolemia, dyslipidemia (e.g., mixed dyslipidemia), orhypertriglyceridemia, comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc fusion proteinsuch as V103 (SEQ ID NO: 11)) at a dose in the range of 100 mg to 600mg, or more specifically at a dose in the range of 100 mg to 300 mg or200 mg to 400 mg, for example, once every day, once every 2, 3, 4, 5, or6 days, once every week, once every 2 weeks, once every 3 weeks, or onceevery 4 weeks (or once every month). In a specific aspect, providedherein is a method of treating, preventing, or managing (e.g.,alleviating one or more symptoms of) hypercholesterolemia, dyslipidemia(e.g., mixed dyslipidemia), or hypertriglyceridemia, comprisingadministering to a subject in need thereof a human FGF21 protein variant(for example, an Fc fusion protein such as V103 (SEQ ID NO: 11)) at adose of approximately 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg,160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg,250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg,340 mg, or 350 mg, for example, once every day, once every 2, 3, 4, 5,or 6 days, once every week, once every 2 weeks, once every 3 weeks, oronce every 4 weeks (or once every month). In a certain aspect, providedherein is a method of treating, preventing, or managing (e.g.,alleviating one or more symptoms of) hypercholesterolemia, dyslipidemia(e.g., mixed dyslipidemia), or hypertriglyceridemia, comprisingadministering to a subject in need thereof a human FGF21 protein variant(for example, an Fc fusion protein such as V103 (SEQ ID NO: 11)) at adose of approximately 300 mg, subcutaneously, once every 4 weeks (oronce every month). In a certain aspect, provided herein is a method oftreating, preventing, or managing (e.g., alleviating one or moresymptoms of) hypercholesterolemia, dyslipidemia (e.g., mixeddyslipidemia), or hypertriglyceridemia, comprising administering to asubject in need thereof a human FGF21 protein variant (for example, anFc fusion protein such as V103 (SEQ ID NO: 11)) at a dose ofapproximately 300 mg, subcutaneously, once every 3 weeks. In a certainaspect, provided herein is a method of treating, preventing, or managing(e.g., alleviating one or more symptoms of) hypercholesterolemia,dyslipidemia (e.g., mixed dyslipidemia), or hypertriglyceridemia,comprising administering to a subject in need thereof a human FGF21protein variant (for example, an Fc fusion protein such as V103 (SEQ IDNO: 11)) at a dose of approximately 300 mg, subcutaneously, once every 2weeks. In a certain aspect, provided herein is a method of treating,preventing, or managing (e.g., alleviating one or more symptoms of)hypercholesterolemia, dyslipidemia (e.g., mixed dyslipidemia), orhypertriglyceridemia, comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc fusion proteinsuch as V103 (SEQ ID NO: 11)) at a dose of approximately 300 mg,subcutaneously, once every week. In a certain aspect, provided herein isa method of treating, preventing, or managing (e.g., alleviating one ormore symptoms of) hypercholesterolemia, dyslipidemia (e.g., mixeddyslipidemia), or hypertriglyceridemia, comprising administering to asubject in need thereof a human FGF21 protein variant (for example, anFc fusion protein such as V103 (SEQ ID NO: 11)) at a dose in the rangeof approximately 200 mg to 250 mg, subcutaneously, once every 3 weeks oronce every 2 weeks. In specific aspects, methods provided herein are fortreating a human subject with hypercholesterolemia (e.g., primaryhypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). Inparticular aspects, methods provided herein are for treating a humansubject with hypertriglyceridemia, in particular, severehypertriglyceridemia. In certain aspects, methods provided herein arefor treating a human subject with type 2 diabetes or a human subject whois obese. In specific aspects, methods provided herein are for treatinga human subject 18 to 55 years of age, who has (i) a BMI within therange of 30-45 kg/m², inclusive (with ethnic adjustment of greater thanor equal to 27.5 kg/m² BMI for Asian subjects), and (ii) triglyceridelevels in the range of 150 to 500 mg/dL (1.69-5.65 mmol/L), prior toadministration of the human FGF21 protein variant.

In specific aspects, provided herein is a method of reducing body weight(e.g., reducing body weight by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, or20%, or more), comprising administering to a subject in need thereof ofa human FGF21 protein variant (for example, an Fc fusion protein such asV103 (SEQ ID NO: 11)) at a dose in the range of 100 mg to 600 mg, ormore specifically at a dose in the range of 200 mg to 400 mg or 100 mgto 300 mg, for example once every day, once every 2, 3, 4, 5, or 6 days,once every week, once every 2 weeks, once every 3 weeks, or once every 4weeks (or once every month). In a certain aspect, provided herein is amethod of reducing body weight (e.g., reducing body weight by at least5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more), comprising administeringto a subject in need thereof of a human FGF21 protein variant (forexample, an Fc fusion protein such as V103 (SEQ ID NO: 11)) at a dose ofapproximately 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg,170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg,260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg,or 350 mg, for example once every day, once every 2, 3, 4, 5, or 6 days,once every week, once every 2 weeks, once every 3 weeks, or once every 4weeks (or once every month). In a certain aspect, provided herein is amethod of reducing body weight (e.g., reducing body weight by at least5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more), comprising administeringto a subject in need thereof of a human FGF21 protein variant (forexample, an Fc fusion protein such as V103 (SEQ ID NO: 11)) at a dose ofapproximately 300 mg, subcutaneously, once every 4 weeks (or once amonth). In a certain aspect, provided herein is a method of reducingbody weight (e.g., reducing body weight by at least 5%, 6%, 7%, 8%, 9%,10%, 15%, or 20% or more), comprising administering to a subject in needthereof of a human FGF21 protein variant (for example, an Fc fusionprotein such as V103 (SEQ ID NO: 11)) at a dose of approximately 300 mg,subcutaneously, once every 3 weeks. In a certain aspect, provided hereinis a method of reducing body weight (e.g., reducing body weight by atleast 5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more), comprisingadministering to a subject in need thereof of a human FGF21 proteinvariant (for example, an Fc fusion protein such as V103 (SEQ ID NO: 11))at a dose of approximately 300 mg, subcutaneously, once every 2 weeks.In a certain aspect, provided herein is a method of reducing body weight(e.g., reducing body weight by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, or20% or more), comprising administering to a subject in need thereof of ahuman FGF21 protein variant (for example, an Fc fusion protein such asV103 (SEQ ID NO: 11)) at a dose of approximately 300 mg, subcutaneously,once every week. In a certain aspect, provided herein is a method ofreducing body weight (e.g., reducing body weight by at least 5%, 6%, 7%,8%, 9%, 10%, 15%, or 20% or more), comprising administering to a subjectin need thereof of a human FGF21 protein variant (for example, an Fcfusion protein such as V103 (SEQ ID NO: 11)) at a dose in the range ofapproximately 200 mg to 250 mg, subcutaneously, once every 3 weeks oronce every 2 weeks. In specific aspects, methods provided herein are fortreating a human subject with hypercholesterolemia (e.g., primaryhypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). Inparticular aspects, methods provided herein are for treating a humansubject with hypertriglyceridemia, in particular, severehypertriglyceridemia. In certain aspects, methods provided herein arefor treating a human subject with type 2 diabetes or a human subject whois obese. In specific aspects, methods provided herein are for treatinga human subject 18 to 55 years of age, who has (i) a BMI within therange of 30-45 kg/m², inclusive (with ethnic adjustment of greater thanor equal to 27.5 kg/m² BMI for Asian subjects), and (ii) triglyceridelevels in the range of 150 to 500 mg/dL (1.69-5.65 mmol/L), at screeningprior to administration of the human FGF21 protein variant.

In particular aspects, provided herein is a method of treating,preventing, or managing (e.g., alleviating one or more symptoms of)obesity, comprising administering to a subject in need thereof a humanFGF21 protein variant (for example, an Fc fusion protein such as V103(SEQ ID NO: 11)) at a dose in the range of 100 mg to 600 mg, or morespecifically at a dose in the range of 200 mg to 400 mg or 100 mg to 300mg, for example once every day, once every 2, 3, 4, 5, or 6 days, onceevery week, once every 2 weeks, once every 3 weeks, or once every 4weeks (or once every month). In a specific aspect, provided herein is amethod of treating, preventing, or managing (e.g., alleviating one ormore symptoms of) obesity, comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc fusion proteinsuch as V103 (SEQ ID NO: 11)) at a dose of approximately 100 mg, 110 mg,120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg,210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg,300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, for example onceevery day, once every 2, 3, 4, 5, or 6 days, once every week, once every2 weeks, once every 3 weeks, or once every 4 weeks (or once everymonth). In a certain aspect, provided herein is a method of treating,preventing, or managing (e.g., alleviating one or more symptoms of)obesity, comprising administering to a subject in need thereof a humanFGF21 protein variant (for example, an Fc fusion protein such as V103(SEQ ID NO: 11)) at a dose of approximately 300 mg, subcutaneously, onceevery 4 weeks (once a month). In a certain aspect, provided herein is amethod of treating, preventing, or managing (e.g., alleviating one ormore symptoms of) obesity, comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc fusion proteinsuch as V103 (SEQ ID NO: 11)) at a dose of approximately 300 mg,subcutaneously, once every 3 weeks. In a certain aspect, provided hereinis a method of treating, preventing, or managing (e.g., alleviating oneor more symptoms of) obesity, comprising administering to a subject inneed thereof a human FGF21 protein variant (for example, an Fc fusionprotein such as V103 (SEQ ID NO: 11)) at a dose of approximately 300 mg,subcutaneously, once every 2 weeks. In a certain aspect, provided hereinis a method of treating, preventing, or managing (e.g., alleviating oneor more symptoms of) obesity, comprising administering to a subject inneed thereof a human FGF21 protein variant (for example, an Fc fusionprotein such as V103 (SEQ ID NO: 11)) at a dose of approximately 300 mg,subcutaneously, once every 1 week. In a certain aspect, provided hereinis a method of treating, preventing, or managing (e.g., alleviating oneor more symptoms of) obesity, comprising administering to a subject inneed thereof a human FGF21 protein variant (for example, an Fc fusionprotein such as V103 (SEQ ID NO: 11)) at a dose in the range ofapproximately 200 mg to 250 mg, subcutaneously, once every 3 weeks oronce every 2 weeks. In specific aspects, methods provided herein are fortreating a human subject with hypercholesterolemia (e.g., primaryhypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). Inparticular aspects, methods provided herein are for treating a humansubject with hypertriglyceridemia, in particular, severehypertriglyceridemia. In certain aspects, methods provided herein arefor treating a human subject with type 2 diabetes or a human subject whois obese. In specific aspects, methods provided herein are for treatinga human subject 18 to 55 years of age, who has (i) a BMI within therange of 30-45 kg/m², inclusive (with ethnic adjustment of greater thanor equal to 27.5 kg/m² BMI for Asian subjects), and (ii) triglyceridelevels in the range of 150 to 500 mg/dL (1.69-5.65 mmol/L), at screeningprior to administration of the human FGF21 protein variant.

In particular aspects, provided herein is a method of treating,preventing, or managing (e.g., alleviating one or more symptoms of)NASH, comprising administering to a subject in need thereof a humanFGF21 protein variant (for example, an Fc fusion protein such as V103(SEQ ID NO: 11)) at a dose in the range of 100 mg to 600 mg, or morespecifically at a dose in the range of 200 mg to 400 mg or 100 mg to 300mg, for example once every day, once every 2, 3, 4, 5, or 6 days, onceevery week, once every 2 weeks, once every 3 weeks, or once every 4weeks (or once every month). In a specific aspect, provided herein is amethod of treating, preventing, or managing (e.g., alleviating one ormore symptoms of) NASH, comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc fusion proteinsuch as V103 (SEQ ID NO: 11)) at a dose of approximately 100 mg, 110 mg,120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg,210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg,300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, for example onceevery day, once every 2, 3, 4, 5, or 6 days, once every week, once every2 weeks, once every 3 weeks, or once every 4 weeks (or once everymonth). In a certain aspect, provided herein is a method of treating,preventing, or managing (e.g., alleviating one or more symptoms of)NASH, comprising administering to a subject in need thereof a humanFGF21 protein variant (for example, an Fc fusion protein such as V103(SEQ ID NO: 11)) at a dose of approximately 300 mg, subcutaneously, onceevery 4 weeks (or once a month). In a certain aspect, provided herein isa method of treating, preventing, or managing (e.g., alleviating one ormore symptoms of) NASH, comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc fusion proteinsuch as V103 (SEQ ID NO: 11)) at a dose of approximately 300 mg,subcutaneously, once every 3 weeks. In a certain aspect, provided hereinis a method of treating, preventing, or managing (e.g., alleviating oneor more symptoms of) NASH, comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc fusion proteinsuch as V103 (SEQ ID NO: 11)) at a dose of approximately 300 mg,subcutaneously, once every 2 weeks. In a certain aspect, provided hereinis a method of treating, preventing, or managing (e.g., alleviating oneor more symptoms of) NASH, comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc fusion proteinsuch as V103 (SEQ ID NO: 11)) at a dose of approximately 300 mg,subcutaneously, once every week. In a certain aspect, provided herein isa method of treating, preventing, or managing (e.g., alleviating one ormore symptoms of) NASH, comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc fusion proteinsuch as V103 (SEQ ID NO: 11)) at a dose in the range of approximately200 mg to 250 mg, subcutaneously, once every 3 weeks or once every 2weeks. In specific aspects, methods provided herein are for treating ahuman subject with hypercholesterolemia (e.g., primaryhypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). Inparticular aspects, methods provided herein are for treating a humansubject with hypertriglyceridemia, in particular, severehypertriglyceridemia. In certain aspects, methods provided herein arefor treating a human subject with type 2 diabetes or a human subject whois obese. In specific aspects, methods provided herein are for treatinga human subject 18 to 55 years of age, who has (i) a BMI within therange of 30-45 kg/m², inclusive (with ethnic adjustment of greater thanor equal to 27.5 kg/m² BMI for Asian subjects), and (ii) triglyceridelevels in the range of 150 to 500 mg/dL (1.69-5.65 mmol/L), prior toadministration of the human FGF21 protein variant.

In particular aspects, provided herein is a method of treating,preventing, or managing (e.g., alleviating one or more symptoms of)NAFLD, comprising administering to a subject in need thereof a humanFGF21 protein variant (for example, an Fc fusion protein such as V103(SEQ ID NO: 11)) at a dose in the range of 100 mg to 600 mg, or morespecifically at a dose in the range of 200 mg to 400 mg or 100 mg to 300mg, for example once every day, once every 2, 3, 4, 5, or 6 days, onceevery week, once every 2 weeks, once every 3 weeks, or once every 4weeks (or once every month). In a specific aspect, provided herein is amethod of treating, preventing, or managing (e.g., alleviating one ormore symptoms of) NAFLD, comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc fusion proteinsuch as V103 (SEQ ID NO: 11)) at a dose of approximately 100 mg, 110 mg,120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg,210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg,300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, for example onceevery day, once every 2, 3, 4, 5, or 6 days, once every week, once every2 weeks, once every 3 weeks, or once every 4 weeks (or once everymonth). In a certain aspect, provided herein is a method of treating,preventing, or managing (e.g., alleviating one or more symptoms of)NAFLD, comprising administering to a subject in need thereof a humanFGF21 protein variant (for example, an Fc fusion protein such as V103(SEQ ID NO: 11)) at a dose of approximately 300 mg, subcutaneously, onceevery 4 weeks (or once a month). In a certain aspect, provided herein isa method of treating, preventing, or managing (e.g., alleviating one ormore symptoms of) NAFLD, comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc fusion proteinsuch as V103 (SEQ ID NO: 11)) at a dose of approximately 300 mg,subcutaneously, once every 3 weeks. In a certain aspect, provided hereinis a method of treating, preventing, or managing (e.g., alleviating oneor more symptoms of) NAFLD, comprising administering to a subject inneed thereof a human FGF21 protein variant (for example, an Fc fusionprotein such as V103 (SEQ ID NO: 11)) at a dose of approximately 300 mg,subcutaneously, once every 2 weeks. In a certain aspect, provided hereinis a method of treating, preventing, or managing (e.g., alleviating oneor more symptoms of) NAFLD, comprising administering to a subject inneed thereof a human FGF21 protein variant (for example, an Fc fusionprotein such as V103 (SEQ ID NO: 11)) at a dose of approximately 300 mg,subcutaneously, once every week. In a certain aspect, provided herein isa method of treating, preventing, or managing (e.g., alleviating one ormore symptoms of) NAFLD, comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc fusion proteinsuch as V103 (SEQ ID NO: 11)) at a dose in the range of approximately200 mg to 250 mg, subcutaneously, once every 3 weeks or once every 2weeks. In specific aspects, methods provided herein are for treating ahuman subject with hypercholesterolemia (e.g., primaryhypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). Inparticular aspects, methods provided herein are for treating a humansubject with hypertriglyceridemia, in particular, severehypertriglyceridemia. In certain aspects, methods provided herein arefor treating a human subject with type 2 diabetes or a human subject whois obese. In specific aspects, methods provided herein are for treatinga human subject 18 to 55 years of age, who has (i) a BMI within therange of 30-45 kg/m², inclusive (with ethnic adjustment of greater thanor equal to 27.5 kg/m² BMI for Asian subjects), and (ii) triglyceridelevels in the range of 150 to 500 mg/dL (1.69-5.65 mmol/L), prior toadministration of the human FGF21 protein variant.

In particular aspects, provided herein is a method of reducing fat orlipids in the liver, comprising administering to a subject in needthereof (e.g., a subject with NASH or NAFLD) a human FGF21 proteinvariant (for example, an Fc fusion protein such as V103 (SEQ ID NO: 11))at a dose in the range of 100 mg to 600 mg, or more specifically at adose in the range of 200 mg to 400 mg or 100 mg to 300 mg, for exampleonce every day, once every 2, 3, 4, 5, or 6 days, once every week, onceevery 2 weeks, once every 3 weeks, or once every 4 weeks (or once everymonth). In a specific aspect, provided herein is a method of reducingfat or lipids in the liver, comprising administering to a subject inneed thereof (e.g., a subject with NASH or NAFLD) a human FGF21 proteinvariant (for example, an Fc fusion protein such as V103 (SEQ ID NO: 11))at a dose of approximately 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330mg, 340 mg, or 350 mg, for example once every day, once every 2, 3, 4,5, or 6 days, once every week, once every 2 weeks, once every 3 weeks,or once every 4 weeks (or once every month). In a certain aspect,provided herein is a method of reducing fat or lipids in the liver,comprising administering to a subject in need thereof (e.g., a subjectwith NASH or NAFLD) a human FGF21 protein variant (for example, an Fcfusion protein such as V103 (SEQ ID NO: 11)) at a dose of approximately300 mg, subcutaneously, once every 4 weeks (or once a month). In acertain aspect, provided herein is a method of reducing fat or lipids inthe liver, comprising administering to a subject in need thereof (e.g.,a subject with NASH or NAFLD) a human FGF21 protein variant (forexample, an Fc fusion protein such as V103 (SEQ ID NO: 11)) at a dose ofapproximately 300 mg, subcutaneously, once every 3 weeks. In a certainaspect, provided herein is a method of reducing fat or lipids in theliver, comprising administering to a subject in need thereof (e.g., asubject with NASH or NAFLD) a human FGF21 protein variant (for example,an Fc fusion protein such as V103 (SEQ ID NO: 11)) at a dose ofapproximately 300 mg, subcutaneously, once every 2 weeks. In a certainaspect, provided herein is a method of reducing fat or lipids in theliver, comprising administering to a subject in need thereof (e.g., asubject with NASH or NAFLD) a human FGF21 protein variant (for example,an Fc fusion protein such as V103 (SEQ ID NO: 11)) at a dose ofapproximately 300 mg, subcutaneously, once every week. In a certainaspect, provided herein is a method of reducing fat or lipids in theliver, comprising administering to a subject in need thereof (e.g., asubject with NASH or NAFLD) a human FGF21 protein variant (for example,an Fc fusion protein such as V103 (SEQ ID NO: 11)) at a dose in therange of approximately 200 mg to 250 mg, subcutaneously, once every 3weeks or once every 2 weeks. In specific aspects, methods providedherein are for treating a human subject with NASH or NAFLD. In specificaspects, methods provided herein are for treating a human subject withhypercholesterolemia (e.g., primary hypercholesterolemia) ordyslipidemia (e.g., mixed dyslipidemia). In particular aspects, methodsprovided herein are for treating a human subject withhypertriglyceridemia, in particular, severe hypertriglyceridemia. Incertain aspects, methods provided herein are for treating a humansubject with type 2 diabetes or a human subject who is obese. Inspecific aspects, methods provided herein are for treating a humansubject 18 to 55 years of age, who has (i) a BMI within the range of30-45 kg/m², inclusive (with ethnic adjustment of greater than or equalto 27.5 kg/m² BMI for Asian subjects), and (ii) triglyceride levels inthe range of 150 to 500 mg/dL (1.69-5.65 mmol/L), at screening prior toadministration of the human FGF21 protein variant.

In specific aspects, provided herein is a method of reducing elevatedLDL cholesterol (LDL-C), total cholesterol (total-C), triglyceride,and/or Apo B, comprising administering to a subject in need thereof ahuman FGF21 protein variant (for example, an Fc fusion protein such asV103 (SEQ ID NO: 11)) at a dose in the range of 100 mg to 600 mg, forexample once every day, once every 2, 3, 4, 5, or 6 days, once everyweek, once every 2 weeks, once every 3 weeks, or once every 4 weeks (oronce every month). In particular aspects, provided herein is a method ofreducing elevated LDL-C, total-C, triglyceride, and/or Apo B, comprisingadministering to a subject in need thereof a human FGF21 protein variant(for example, an Fc fusion protein such as V103 (SEQ ID NO: 11)) at adose in the range of 100 mg to 600 mg, or more specifically at a dose inthe range of 200 mg to 400 mg or 100 mg to 300 mg. In a specific aspect,provided herein is a method of reducing elevated LDL-C, total-C,triglyceride, and/or Apo B, comprising administering to a subject inneed thereof a human FGF21 protein variant (for example, an Fc fusionprotein such as V103 (SEQ ID NO: 11)) at a dose of approximately 100 mg,110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg,200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg,290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, for exampleonce every day, once every 2, 3, 4, 5, or 6 days, once every week, onceevery 2 weeks, once every 3 weeks, or once every 4 weeks (or once everymonth). In a certain aspect, provided herein is a method of reducingelevated LDL-C, total-C, triglyceride, and/or Apo B, comprisingadministering to a subject in need thereof a human FGF21 protein variant(for example, an Fc fusion protein such as V103 (SEQ ID NO: 11)) at adose of approximately 300 mg, subcutaneously, once every 4 weeks (oronce a month). In a certain aspect, provided herein is a method ofreducing elevated LDL-C, total-C, triglyceride, and/or Apo B, comprisingadministering to a subject in need thereof a human FGF21 protein variant(for example, an Fc fusion protein such as V103 (SEQ ID NO: 11)) at adose of approximately 300 mg, subcutaneously, once every 3 weeks. In acertain aspect, provided herein is a method of reducing elevated LDL-C,total-C, triglyceride, and/or Apo B, comprising administering to asubject in need thereof a human FGF21 protein variant (for example, anFc fusion protein such as V103 (SEQ ID NO: 11)) at a dose ofapproximately 300 mg, subcutaneously, once every 2 weeks. In a certainaspect, provided herein is a method of reducing elevated LDL-C, total-C,triglyceride, and/or Apo B, comprising administering to a subject inneed thereof a human FGF21 protein variant (for example, an Fc fusionprotein such as V103 (SEQ ID NO: 11)) at a dose of approximately 300 mg,subcutaneously, once every week. In a certain aspect, provided herein isa method of reducing elevated LDL-C, total-C, triglyceride, and/or ApoB, comprising administering to a subject in need thereof a human FGF21protein variant (for example, an Fc fusion protein such as V103 (SEQ IDNO: 11)) at a dose in the range of approximately 200 mg to 250 mg,subcutaneously, once every 3 weeks or once every 2 weeks. In a specificaspect, methods provided herein reduce elevated LDL-C, total-C,triglyceride, and/or Apo B by at least about 5%, 6%, 7%, 8%, 9%, 10%,11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65%, 70%, 75%, 80%, 85%, or 90% or more. In specific aspects, methodsprovided herein are for treating a human subject withhypercholesterolemia (e.g., primary hypercholesterolemia) ordyslipidemia (e.g., mixed dyslipidemia). In particular aspects, methodsprovided herein are for treating a human subject withhypertriglyceridemia, in particular, severe hypertriglyceridemia. Incertain aspects, methods provided herein are for treating a humansubject with type 2 diabetes or a human subject who is obese. Inspecific aspects, methods provided herein are for treating a humansubject 18 to 55 years of age, who has (i) a BMI within the range of30-45 kg/m², inclusive (with ethnic adjustment of greater than or equalto 27.5 kg/m² BMI for Asian subjects), and (ii) triglyceride levels inthe range of 150 to 500 mg/dL (1.69-5.65 mmol/L), at screening prior toadministration of the human FGF21 protein variant.

In specific aspects, provided herein is a method of increasing HDL-C(e.g., increasing HDL-C by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%,40%, 45%, or 50%), comprising administering to a subject in need thereofa therapeutically effective amount of a fusion protein comprising ahuman FGF21 protein variant (for example, an Fc fusion protein such asV103 (SEQ ID NO: 11)) at a dose in the range of 100 mg to 600 mg, forexample once every day, once every 2, 3, 4, 5, or 6 days, once everyweek, once every 2 weeks, once every 3 weeks, or once every 4 weeks (oronce every month). In particular aspects, provided herein is a method ofincreasing HDL-C (e.g., increasing HDL-C by at least 5%, 10%, 15%, 20%,25%, 30%, 35%, 40%, 45%, or 50%), comprising administering to a subjectin need thereof a human FGF21 protein variant (for example, an Fc fusionprotein such as V103 (SEQ ID NO: 11)) at a dose in the range of about200 mg to 400 mg. In a specific aspect, provided herein is a method ofincreasing HDL-C (e.g., increasing HDL-C by at least 5%, 10%, 15%, 20%,25%, 30%, 35%, 40%, 45%, or 50%), comprising administering to a subjectin need thereof a human FGF21 protein variant (for example, an Fc fusionprotein such as V103 (SEQ ID NO: 11)) at a dose of approximately 100 mg,110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg,200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg,290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, for exampleonce every day, once every 2, 3, 4, 5, or 6 days, once every week, onceevery 2 weeks, once every 3 weeks, or once every 4 weeks (or once everymonth). In a certain aspect, provided herein is a method of increasingHDL-C (e.g., increasing HDL-C by at least 5%, 10%, 15%, 20%, 25%, 30%,35%, 40%, 45%, or 50%), comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc fusion proteinsuch as V103 (SEQ ID NO: 11)) at a dose of approximately 300 mg,subcutaneously, once every 4 weeks (or once a month) or once every 3weeks or once every 2 weeks. In a certain aspect, provided herein is amethod of increasing HDL-C (e.g., increasing HDL-C by at least 5%, 10%,15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%), comprising administering toa subject in need thereof a human FGF21 protein variant (for example, anFc fusion protein such as V103 (SEQ ID NO: 11)) at a dose in the rangeof approximately 200 mg to 250 mg, subcutaneously, once every 3 weeks oronce every 2 weeks. In specific aspects, methods provided herein are fortreating a human subject with hypercholesterolemia (e.g., primaryhypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). Inparticular aspects, methods provided herein are for treating a humansubject with hypertriglyceridemia, in particular, severehypertriglyceridemia. In certain aspects, methods provided herein arefor treating a human subject with type 2 diabetes or a human subject whois obese. In specific aspects, methods provided herein are for treatinga human subject 18 to 55 years of age, who has (i) a BMI within therange of 30-45 kg/m², inclusive (with ethnic adjustment of greater thanor equal to 27.5 kg/m² BMI for Asian subjects), and (ii) triglyceridelevels in the range of 150 to 500 mg/dL (1.69-5.65 mmol/L), at screeningprior to administration of the human FGF21 protein variant.

In particular aspects, provided herein is a method of reducingtriglyceride levels, for example fasting triglyceride levels, (e.g.,reducing blood/serum triglyceride levels by at least 20%, 30%, 40%, 50%,60%, 70%, 80%, or 90% or more), comprising administering to a subject inneed thereof a human FGF21 protein variant (for example, an Fc fusionprotein such as V103 (SEQ ID NO: 11)) at a dose in the range of 100 mgto 600 mg, for example once every day, once every 2, 3, 4, 5, or 6 days,once every week, once every 2 weeks, once every 3 weeks, or once every 4weeks (or once every month). In particular aspects, provided herein is amethod of reducing triglyceride levels (e.g., reducing blood/serumtriglyceride levels by at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, or90% or more), comprising administering to a subject in need thereof ahuman FGF21 protein variant (for example, an Fc fusion protein such asV103 (SEQ ID NO: 11)) at a dose in the range of about 200 mg to 400 mg.In a specific aspect, provided herein is a method of reducingtriglyceride levels, for example fasting triglyceride levels, (e.g.,reducing blood/serum triglyceride levels by at least 20%, 30%, 40%, 50%,60%, 70%, 80%, or 90% or more), comprising administering to a subject inneed thereof a human FGF21 protein variant (for example, an Fc fusionprotein such as V103 (SEQ ID NO: 11)) at a dose of approximately 100 mg,110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg,200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg,290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, for exampleonce every day, once every 2, 3, 4, 5, or 6 days, once every week, onceevery 2 weeks, once every 3 weeks, or once every 4 weeks (or once everymonth). In a certain aspect, provided herein is a method of reducingtriglyceride levels, for example fasting triglyceride levels, (e.g.,reducing blood/serum triglyceride levels by at least 20%, 30%, 40%, 50%,60%, 70%, 80%, or 90% or more), comprising administering to a subject inneed thereof a human FGF21 protein variant (for example, an Fc fusionprotein such as V103 (SEQ ID NO: 11)) at a dose of approximately 300 mg,subcutaneously, once every 4 weeks (or once a month), or once every 3weeks or once every 2 weeks. In a certain aspect, provided herein is amethod of reducing triglyceride levels, for example fasting triglyceridelevels, (e.g., reducing blood/serum triglyceride levels by at least 20%,30%, 40%, 50%, 60%, 70%, 80%, or 90% or more), comprising administeringto a subject in need thereof a human FGF21 protein variant (for example,an Fc fusion protein such as V103 (SEQ ID NO: 11)) at a dose in therange of approximately 200 mg to 250 mg, subcutaneously, once every 3weeks or once every 2 weeks or once every week. In a specific aspect,methods provided herein are able to reduce triglyceride levels, forexample fasting triglyceride levels, by at least about 40% or at leastabout 50% for a period of at least 1-4 weeks. In a certain aspect,provided herein is a method of reducing triglyceride levels, for examplefasting triglyceride levels, by at least about 40% to 60% (e.g., atleast about 50%) for a period of at least 1-4 weeks, comprisingadministering to a subject in need thereof a human FGF21 protein variant(for example, an Fc fusion protein such as V103 (SEQ ID NO: 11)) at adose of approximately 300 mg, subcutaneously, once every 4 weeks (oronce a month) or once every 3 weeks or once every 2 weeks. In a certainaspect, provided herein is a method of reducing triglyceride levels, forexample fasting triglyceride levels, by at least about 40% to 60% (e.g.,at least about 50%) for a period of at least 1-4 weeks, comprisingadministering to a subject in need thereof a human FGF21 protein variant(for example, an Fc fusion protein such as V103 (SEQ ID NO: 11)) at adose of in the range of 200 mg to 300 mg (e.g., at least 220 mg, 230 mg,240 mg, 250 mg, 260 mg, 270 mg, or 280 mg), subcutaneously, once every 4weeks (or once a month) or once every 3 weeks or once every 2 weeks. Inspecific aspects, methods provided herein are for treating a humansubject with hypercholesterolemia (e.g., primary hypercholesterolemia)or dyslipidemia (e.g., mixed dyslipidemia). In particular aspects,methods provided herein are for treating a human subject withhypertriglyceridemia, in particular, severe hypertriglyceridemia. Incertain aspects, methods provided herein are for treating a humansubject with type 2 diabetes or a human subject who is obese. Inspecific aspects, methods provided herein are for treating a humansubject 18 to 55 years of age, who has (i) a BMI within the range of30-45 kg/m², inclusive (with ethnic adjustment of greater than or equalto 27.5 kg/m² BMI for Asian subjects), and (ii) triglyceride levels inthe range of 150 to 500 mg/dL (1.69-5.65 mmol/L), at screening prior toadministration of the human FGF21 protein variant.

In a specific aspect, methods provided herein are able to reducetriglyceride levels, for example fasting triglyceride levels, by atleast about 40% or at least about 50%, for example for a period of atleast 1-4 weeks, said method comprising administering to a subject inneed thereof a human FGF21 protein variant (for example, an Fc fusionprotein such as V103 (SEQ ID NO: 11)) at a dose in the range ofapproximately 250 mg to 300 mg (e.g., 300 mg), subcutaneously, onceevery 4 weeks (or once a month) or once every 3 weeks. In a certainaspect, provided herein is a method of reducing triglyceride levels, forexample fasting triglyceride levels, by at least about 40% to 60% (e.g.,at least about 50%), for example for a period of at least 1-4 weeks,said method comprising administering to a subject in need thereof ahuman FGF21 protein variant (for example, an Fc fusion protein such asV103 (SEQ ID NO: 11)) at a dose in the range of approximately 250 mg to300 mg (e.g., 300 mg), subcutaneously, once every 4 weeks (or once amonth) or once every 3 weeks or once every 2 weeks.

In particular aspects, provided herein is a method of reducingcardiovascular risk (e.g., by at least about 5%, 10%, 15%, 20%, 25%,30%, 35%, 40%, 45%, or 50% or more), comprising administering to asubject in need thereof a therapeutically effective amount of a humanFGF21 protein variant (for example, an Fc fusion protein such as V103(SEQ ID NO: 11)) at a dose in the range of 100 mg to 600 mg, for exampleonce every day, once every 2, 3, 4, 5, or 6 days, once every week, onceevery 2 weeks, once every 3 weeks, or once every 4 weeks (or once everymonth). In particular aspects, provided herein is a method of reducingcardiovascular risk (e.g., by at least about 5%, 10%, 15%, 20%, 25%,30%, 35%, 40%, 45%, or 50% or more), comprising administering to asubject in need thereof a human FGF21 protein variant (for example, anFc fusion protein such as V103 (SEQ ID NO: 11)) at a dose in the rangeof about 200 mg to 400 mg or 100 mg to 300 mg, for example once a week,once every 2 weeks, once every 3 weeks, or once every 4 weeks. In aspecific aspect, provided herein is a method of reducing cardiovascularrisk (e.g., by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%,45%, or 50% or more), comprising administering to a subject in needthereof a human FGF21 protein variant (for example, an Fc fusion proteinsuch as V103 (SEQ ID NO: 11)) at a dose of approximately 100 mg, 110 mg,120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg,210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg,300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, for example onceevery day, once every 2, 3, 4, 5, or 6 days, once every week, once every2 weeks, once every 3 weeks, or once every 4 weeks (or once everymonth). In a certain aspect, provided herein is a method of reducingcardiovascular risk (e.g., by at least about 5%, 10%, 15%, 20%, 25%,30%, 35%, 40%, 45%, or 50% or more), comprising administering to asubject in need thereof a human FGF21 protein variant (for example, anFc fusion protein such as V103 (SEQ ID NO: 11)) at a dose ofapproximately 300 mg, subcutaneously, once every 4 weeks (or once everymonth). In a certain aspect, provided herein is a method of reducingcardiovascular risk (e.g., by at least about 5%, 10%, 15%, 20%, 25%,30%, 35%, 40%, 45%, or 50% or more), comprising administering to asubject in need thereof a human FGF21 protein variant (for example, anFc fusion protein such as V103 (SEQ ID NO: 11)) at a dose ofapproximately 300 mg, subcutaneously, once every 3 weeks. In a certainaspect, provided herein is a method of reducing cardiovascular risk(e.g., by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or50%), comprising administering to a subject in need thereof a humanFGF21 protein variant (for example, an Fc fusion protein such as V103(SEQ ID NO: 11)) at a dose in the range of approximately 200 mg to 250mg, subcutaneously, every 3 weeks. In a certain aspect, provided hereinis a method of reducing cardiovascular risk (e.g., by at least about 5%,10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% or more), comprisingadministering to a subject in need thereof a human FGF21 protein variant(for example, an Fc fusion protein such as V103 (SEQ ID NO: 11)) at adose of approximately 300 mg, subcutaneously, once every 2 weeks or onceevery week. In specific aspects, methods provided herein are fortreating a human subject with hypercholesterolemia (e.g., primaryhypercholesterolemia) or dyslipidemia (e.g., mixed dyslipidemia). Inparticular aspects, methods provided herein are for treating a humansubject with hypertriglyceridemia, in particular, severehypertriglyceridemia. In certain aspects, methods provided herein arefor treating a human subject with type 2 diabetes or a human subject whois obese. In specific aspects, methods provided herein are for treatinga human subject 18 to 55 years of age, who has (i) a BMI within therange of 30-45 kg/m², inclusive (with ethnic adjustment of greater thanor equal to 27.5 kg/m² BMI for Asian subjects), and (ii) triglyceridelevels in the range of 150 to 500 mg/dL (1.69-5.65 mmol/L), prior toadministration of the human FGF21 protein variant.

In specific aspects, methods provided herein are for treating a humansubject 18 to 55 years of age, who has (i) a BMI within the range of30-45 kg/m², inclusive (with ethnic adjustment of greater than or equalto 27.5 kg/m² BMI for Asian subjects), and (ii) triglyceride levels inthe range of 150 to 500 mg/dL (1.69-5.65 mmol/L), at screening prior toadministration of the human FGF21 protein variant.

In various aspects, methods provided herein are for treating a humansubject at least 18 years of age. In other aspects, methods providedherein are for treating a human subject at least 55 years of age. Inother aspects, methods provided herein are for treating a human subjectat least 60 years of age. In other aspects, methods provided herein arefor treating a human subject at least 65 years of age.

In specific aspects, methods provided herein are for treating a humansubject who has a BMI within the range of 30 to 45 kg/m². In certainaspects, methods provided herein are for treating a human subject whohas a BMI greater than or equal to 25 kg/m². In certain aspects, methodsprovided herein are for treating a human subject who has a BMI greaterthan or equal to 30 kg/m². In certain aspects, methods provided hereinare for treating a human subject who has a BMI greater than or equal to35 kg/m². In some aspects, methods provided herein are for treating ahuman subject with a BMI (with ethnic adjustment)≥27.5 for Asianindividuals.

In particular aspects, methods provided herein are for treating a humansubject who has triglyceride levels in the range of 150-500 mg/dL(1.69-5.65 mmol/L), prior to administration of a human FGF21 proteinvariant. In particular aspects, methods provided herein are for treatinga human subject who has triglyceride levels that is at least 150 mg/dL(at least 1.69 mmol/L), prior to administration of a human FGF21 proteinvariant. In particular aspects, methods provided herein are for treatinga human subject who has triglyceride levels that is at least 200 mg/dL,at screening prior to administration of a human FGF21 protein variant.In particular aspects, methods provided herein are for treating a humansubject who has triglyceride levels that is at least 500 mg/dL (at least5.65 mmol/L), prior to administration of a human FGF21 protein variant.

In certain aspects, human subject with one or more of the following arenot treated according to the methods provided herein:

-   -   History of hepatobilliary disease, cholelithiasis, or biliary        sludge by history or at screening ultrasound, hepatic        encephalopathy, esophageal varices, or porticaval shunt;    -   Liver disease or liver injury as indicated by abnormal liver        function tests (ALT, AST, GGT, alkaline phosphatase, or serum        bilirubin) above upper limit of normal at screening or baseline;    -   Chronic infection with Human Immunodeficiency Virus (HIV),        Hepatitis B (HBV) or Hepatitis C (HCV);    -   A positive HBV surface antigen (HBsAg) test, or if standard        local practice, a positive HBV core antigen test.    -   Positive (detectable) HCV RNA;    -   History of pancreatic injury or pancreatitis, or other        pancreatic disease;    -   Amylase or lipase above ULN at screening or baseline;    -   History of hypersensitivity to drugs of similar biological        class, FGF21 protein analogue, or Fc fusion proteins;    -   History of bone disorders including but not limited to        osteoporosis, osteopenia, osteomalacia, severe vitamin D        deficiency; and    -   Plasma 25-hydroxyvitamin D level is below the lower limit of the        normal range at screening.

In one aspect, provided herein are combination therapies for treating,preventing, or managing a metabolic disorder or a cardiovasculardisorder comprising administering a therapeutically effective amount ofan FGF21 protein variant described herein (e.g., Fc fusion protein suchas V103) and one or more additional therapeutically active agents (e.g.,therapeutic agents for metabolic disorders or cardiovascular disorders).Non-limiting examples of other therapeutically active agents for use incombination with FGF21 protein variants provided herewith includeobesity therapies (e.g., phentermine/topiramate, orlistat, lorcaserin,liraglutide, bupropion/naltrexone), high blood pressure therapies (e.g.,diuretics, beta-blockers, alpha-blockers, ACE inhibitors, Angiotensin IIReceptor Blockers (ARBs), direct renin inhibitors, calcium channelblockers, central agonists, peripheral adrenergic blockers,vasodialators, and combinations), diabetic therapies (e.g., insulin,alpha-glucosidase inhibitors, biguanides, dopamine agonist, DPP-4inhibitors, glucagon-like peptides, meglitinides, sodium glucosetransporter (SGLT) inhibitors, sulfonylureas, thiazolidinediones,amylinomimetics), NAFLD/NASH and cardiovascular therapies (e.g.,statins, fibrates, aspirin, anticoagulants).

In one aspect, provided herein are combination therapies for treating,preventing, or managing a metabolic disorder or a cardiovasculardisorder comprising administering a therapeutically effective amount ofan FGF21 protein variant described herein (e.g., Fc fusion protein suchas V103) and one or more therapeutically active agents selected from thefollowing: amiloride (Midamor), bumetanide (Bumex), chlorthalidone(Hygroton), chlorothiazide (Diuril), furosemide (Lasix),hydrochlorothiazide or HCTZ (Esidrix, Hydrodiuril, Microzide),indapamide (Lozol), metolazone (Mykrox, Zaroxolyn), spironolactone(Aldactone), triamterene (Dyrenium), Acebutolol (Sectral), Atenolol(Tenormin), Betaxolol (Kerlone), Bisoprolol (Zebeta), Carteolol(Cartrol), Metoprolol (Lopressor, Toprol XL), Nadolol (Corgard),Nebivolol (Bystolic), Penbutolol (Levatol), Pindolol (Visken),Propranolol (Inderal), Sotalol (Betapace), Timolol (Blocadren),Doxazosin (Cardura), Prazosin (Minipress), Terazosin (Hytrin),Benazepril (Lotensin), Captopril (Capoten), Enalapril (Vasotec),Fosinopril (Monopril), Lisinopril (Prinivil, Zestril), Moexipril(Univasc), Perindopril (Aceon), Quinapril (Accupril), Ramipril (Altace),Trandolapril (Mavik), Norvasc (amlodipine), Plendil (felodipine),DynaCirc (isradipine), Cardene (nicardipine), Procardia XL, Adalat(nifedipine), Cardizem, Dilacor, Tiazac, Diltia XL (diltiazem), Sular(Nisoldipine), Isoptin, Calan, Verelan, Covera-HS (verapamil), Capoten(captopril), Vasotec (enalapril), Prinivil, Zestril (lisinopril),Lotensin (benazepril), Monopril (fosinopril), Altace (ramipril),Accupril (quinapril), Aceon (perindopril), Mavik (trandolapril), Univasc(moexipril), Atacand (candesartan), Avapro (irbesartan), Benicar(olmesartan), Cozaar (losartan), Diovan (valsartan), Micardis(telmisartan), Teveten (eprosartan), Chlorthalidone (Hygroton),Chlorothiazide (Diuril), Hydrochlorothiazide or HCTZ (Esidrix,Hydrodiuril, Microzide), Indapamide (Lozol), Metolazone (Mykrox,Zaroxolyn), Amiloride (Midamor), Bumetanide (Bumex), Furosemide (Lasix),Spironolactone (Aldactone), Triamterene (Dyrenium), Acebutolol(Sectral), Atenolol (Tenormin), Betaxolol (Kerlone), Bisoprolol (Zebeta,Ziac), Carteolol (Cartrol), Carvedilol (Coreg), Labetalol (Normodyne,Trandate), Metoprolol (Lopressor, Toprol-XL), Nadolol (Corgard),Nebivolol (Bystolic), Penbutolol (Levatol), Pindolol (Visken),Propanolol (Inderal), Sotalol (Betapace), Timolol (Blocadren), fibricacid derivatives, niacin, and omega-3 fatty acids, fenofibrate,gemfibrozil, atorvastatin, fluvastatin, lovastatin, pitavastatin,pravastatin, rosuvastatin, simvastatin, pramlintide, acarbose (Precose),miglitol (Glyset), metformin, bromocriptine, alogliptin, linagliptin,saxagliptin, sitagliptin, albiglutide (Tanzeum), dulaglutide(Trulicity), exenatide (Byetta), exenatide extended-release (Bydureon),liraglutide (Victoza), nateglinide (Starlix), repaglinide (Prandin),repaglinide-metformin (Prandimet), dapagliflozin (Farxiga),dapagliflozin-metformin (Xigduo XR), canagliflozin (Invokana),canagliflozin-metformin (Invokamet), empagliflozin (Jardiance),empagliflozin-linagliptin (Glyxambi), empagliflozin-metformin(Synjardy), sotagliflozin, tofogliflozin, remogliflozin, luseogliflozin,ipragliflozin, atigliflozin, bexagliflozin, henagliflozin,licogliflozin, glimepiride (Amaryl), glimepiride-pioglitazone (Duetact),glimeperide-rosiglitazone (Avandaryl), gliclazide, glipizide-metformin(Metaglip), glyburide (DiaBeta, Glynase, Micronase), glyburide-metformin(Glucovance), chlorpropamide (Diabinese), tolazamide (Tolinase),tolbutamide (Orinase, Tol-Tab), rosiglitazone (Avandia),rosiglitazone-glimepiride (Avandaryl), rosiglitizone-metformin (AmarylM), pioglitazone (Actos), pioglitazone-alogliptin (Oseni),pioglitazone-glimepiride (Duetact), and pioglitazone-metformin (ActoplusMet, Actoplus Met XR).

In some embodiments, the sodium glucose transporter (SGLT) inhibitor isselected from dapagliflozin, empagliflozin, canagliflozin,ertugliflozin, sotagliflozin, tofogliflozin, remogliflozin,luseogliflozin, ipragliflozin, atigliflozin, bexagliflozin,henagliflozin, licogliflozin, and a pharmaceutically acceptable salt ofany of these. In some embodiments, the sodium glucose transporter (SGLT)inhibitor is dapagliflozin. In some embodiments, the sodium glucosetransporter (SGLT) inhibitor is empagliflozin. In some embodiments, thesodium glucose transporter (SGLT) inhibitor is canagliflozin. In someembodiments, the sodium glucose transporter (SGLT) inhibitor isertugliflozin. In some embodiments, the sodium glucose transporter(SGLT) inhibitor is licogliflozin.

In some embodiments, the sodium glucose transporter (SGLT) inhibitor isdapagliflozin or a pharmaceutically acceptable salt thereof. In someembodiments, the sodium glucose transporter (SGLT) inhibitor isempagliflozin or a pharmaceutically acceptable salt thereof. In someembodiments, the sodium glucose transporter (SGLT) inhibitor iscanagliflozin or a pharmaceutically acceptable salt thereof. In someembodiments, the sodium glucose transporter (SGLT) inhibitor isertugliflozin or a pharmaceutically acceptable salt thereof. In someembodiments, the sodium glucose transporter (SGLT) inhibitor islicogliflozin or a pharmaceutically acceptable salt thereof.

In specific aspects, methods provided herein comprising administering anFGF21 protein variant (e.g., Fc fusion protein such as V103) are for useas an adjunct to diet (e.g., healthy diet, calorie restricted diet),exercise, and/or other lifestyle modifications.

FGF21 Variants

Modifications to FGF21 can improve the half-life and/or potency overwild-type FGF21 to provide an FGF21 therapeutic for treating or managing(e.g., alleviating one or more symptoms of a disorder) metabolic orcardiovascular disorders or for reducing cardiovascular risk, withclinically preferred dosing regimens.

In specific aspects, provided herein are FGF21 protein variants, forexample FGF21 protein variants described in Table 1, for use in themethods for treating or managing (e.g., alleviating one or more symptomsof a disorder) metabolic or cardiovascular disorders or for reducingcardiovascular risk.

Acceptable amino acid substitutions and modifications which constitutedifferences between the FGF21 polypeptide and protein variants andmutants of the methods provided herein and wild-type FGF21 include, butare not limited to, one or more amino acid substitutions, includingsubstitutions with non-naturally occurring amino acid analogs, andtruncations. Thus, FGF21 protein variants include, but are not limitedto, site-directed FGF21 mutants, truncated FGF21 polypeptides,proteolysis-resistant FGF21 mutants, aggregation-reducing FGF21 mutants,FGF21 combination mutants, FGF21 conjugates (e.g., fatty acid-FGF21conjugate, PEG-FGF21 conjugate) and FGF21 fusion proteins (e.g., Fcdomain fusion protein, human serum albumin fusion protein).

In one aspect, an FGF21 protein variant comprises substitutions at oneor more or all of the following residues made relative to wild typeFGF21 according to the numbering scheme of SEQ ID NO: 1: Q55, R105,G148, K150, P158, S195, P199, and G202.

In one aspect, an FGF21 protein variant comprises one or more or all ofthe following substitutions made relative to wild type FGF21 accordingto the numbering scheme of SEQ ID NO: 1: Q55C, R105K, G148C, K150R,P158S, S195A, P199G, and G202A.

Exemplary fusion protein sequences provided herein are listed inTable 1. The descriptions of said fusions include the FGF21 variant and,where applicable, a linker. The changes or substitutions employed by theFGF21 variant are numbered and described relative to wild-type FGF21.

By way of example, “Variant 101 (V101)” (SEQ ID NO:10) is an Fc-FGF21fusion with a two amino acid linker and the following substitutions maderelative to wild type FGF21 according to the numbering scheme of SEQ IDNO: 1: Q55C, A109T, G148C, K150R, P158S, P174L, S195A, P199G, G202A.

By way of another example, “Variant 103 (V103)” (SEQ ID NO:11) is anFc-FGF21 fusion with a two amino acid linker (GS) and the followingsubstitutions made relative to wild type FGF21 according to thenumbering scheme of SEQ ID NO: 1: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, G202A.

TABLE 1  Exemplary FGF21 Variants SEQ ID NO: Sequence Description* 7DKTHTCPPCP APEAAGGPSV FLFPPKPKDT Full Length N-termLMISRTPEVT CVVVDVSHED PEVKFNWYVD Fc-Fusion with 2 AAGVEVHNAKTK PREEQYNSTY RVVSVLTVLH Linker (GS) and WTQDWLNGKEYK CKVSNKALPA PIEKTISKAK FGF21 GQPREPQVYT LPPSREEMTK NQVSLTCLVKGFYPSDIAVE WESNGQPENN YKTTPPVLDS DGSFFLYSKL TVDKSRWQQG NVFSCSVMHEALHNHYTQKS LSLSPGKGSD SSPLLQFGGQ VRQRYLYTDD AQQTEAHLEI REDGTVGGAADQSPESLLQL KALKPGVIQI LGVKTSRFLC QRPDGALYGS LHFDPEACSF RELLLEDGYNVYQSEAHGLP LHLPGNKSPH RDPAPRGPAR FLPLPGLPPA LPEPPGILAP QPPDVGSSDPLSMVGPSQGR SPSYAS 8 DKTHTCPPCP APEAAGGPSV FLFPPKPKDT Full Length N-termLMISRTPEVT CVVVDVSHED PEVKFNWYVD Fc-Fusion with 15 AAGVEVHNAKTK PREEQYNSTY RVVSVLTVLH Linker (GGGGS x 3)QDWLNGKEYK CKVSNKALPA PIEKTISKAK between Fc and WTGQPREPQVYT LPPSREEMTK NQVSLTCLVK FGF21 GFYPSDIAVE WESNGQPENN YKTTPPVLDSDGSFFLYSKL TVDKSRWQQG NVFSCSVMHE ALHNHYTQKS LSLSPGKGGG GSGGGGSGGGGSDSSPLLQF GGQVRQRYLY TDDAQQTEAH LEIREDGTVG GAADQSPESL LQLKALKPGVIQILGVKTSR FLCQRPDGAL YGSLHFDPEA CSFRELLLED GYNVYQSEAH GLPLHLPGNKSPHRDPAPRG RARFLPLPGL PPALPEPPGI LAPQPPDVGS SDPLSMVGPS QGRSPSYAS 9DSSPLLQFGG QVRQRYLYTD DAQETEAHLE Variant #76 (V76) =IREDGTVGGA AHQSPESLLE LKALKPGVIQ Protein with 9 totalILGVKTSRFL CQKPDGALYG SLHFDPEACS mutations relative toFRELLLEDGY NVYQSEAHGL PLHLPGNRSP wild-type FGF21 (asHCDPAPQGPA RFLPLPGLPP ALPEPPGILA in W001/018172)PQPPDVGSSD PLAMVGPSQG RSPSYAS 10 DKTHTCPPCP APEAAGGPSV FLFPPKPKDTVariant #101 (V101) = LMISRTPEVT CVVVDVSHED PEVKFNWYVD N-term Fc FusionGVEVHNAKTK PREEQYNSTY RVVSVLTVLH with the 2 AA linkerQDWLNGKEYK CKVSNKALPA PIEKTISKAK (GS) between Fc andGQPREPQVYT LPPSREEMTK NQVSLTCLVK FGF21 = (Q55C, A109T,GFYPSDIAVE WESNGQPENN YKTTPPVLDS G148C, K150R, P158S,DGSFFLYSKL TVDKSRWQQG NVFSCSVMHE 5195A, P199G, G202A)ALHNHYTQKS LSLSPGKGSD SSPLLQFGGQ VRQRYLYTDD ACQTEAHLEI REDGTVGGAADQSPESLLQL KALKPGVIQI LGVKTSRFLC QRPDGTLYGS LHFDPEACSF RELLLEDGYNVYQSEAHGLP LHLPCNRSPH RDPASRGPAR FLPLPGLPPA LPEPPGILAP QPPDVGSSDPLAMVGGSQAR SPSYAS 11 DKTHTCPPCP APEAAGGPSV FLFPPKPKDTVariant #103 (V103) = LMISRTPEVT CVVVDVSHED PEVKFNWYVD N-term Fc FusionGVEVHNAKTK PREEQYNSTY RVVSVLTVLH with the 2 AA linkerQDWLNGKEYK CKVSNKALPA PIEKTISKAK (GS) =(Q55C, R105K,GQPREPQVYT LPPSREEMTK NQVSLTCLVK G148C, K150R, P158S,GFYPSDIAVE WESNGQPENN YKTTPPVLDS S195A, P199G, G202A)DGSFFLYSKL TVDKSRWQQG NVFSCSVMHE ALHNHYTQKS LSLSPGKGSD SSPLLQFGGQVRQRYLYTDD ACQTEAHLEI REDGTVGGAA DQSPESLLQL KALKPGVIQI LGVKTSRFLCQKPDGALYGS LHFDPEACSF RELLLEDGYN VYQSEAHGLP LHLPCNRSPH RDPASRGPARFLPLPGLPPA LPEPPGILAP QPPDVGSSDP LAMVGGSQAR SPSYAS 12DKTHTCPPCP APEAAGGPSV FLFPPKPKDT Variant #188 = V103LMISRTPEVT CVVVDVSHED PEVKFNWYVD with 15 AA LinkerGVEVHNAKTK PREEQYNSTY RVVSVLTVLH (GGGGS x 3) betweenQDWLNGKEYK CKVSNKALPA PIEKTISKAK Fc and FGF21GQPREPQVYT LPPSREEMTK NQVSLTCLVK =(Q55C, R105K,GFYPSDIAVE WESNGQPENN YKTTPPVLDS G148C, K150R, P158S,DGSFFLYSKL TVDKSRWQQG NVFSCSVMHE S195A, P199G, G202A)ALHNHYTQKS LSLSPGKGGG GSGGGGSGGG GSDSSPLLQF GGQVRQRYLY TDDACQTEAHLEIREDGTVG GAADQSPESL LQLKALKPGV IQILGVKTSR FLCQKPDGAL YGSLHFDPEACSFRELLLED GYNVYQSEAH GLPLHLPCNR SPHRDPASRG RARFLPLPGL PPALPEPPGILAPQPPDVGS SDPLAMVGGS QARSPSYAS 13 DKTHTCPPCP APEAAGGPSV FLFPPKPKDTVariant #204 = V101 LMISRTPEVT CVVVDVSHED PEVKFNWYVD with 15 AA LinkerGVEVHNAKTK PREEQYNSTY RVVSVLTVLH (GGGGS x 3) betweenQDWLNGKEYK CKVSNKALPA PIEKTISKAK Fc and FGF21 = (Q55C,GQPREPQVYT LPPSREEMTK NQVSLTCLVK A109T, G148C, K150R,GFYPSDIAVE WESNGQPENN YKTTPPVLDS P158S, S195A, P199G,DGSFFLYSKL TVDKSRWQQG NVFSCSVMHE G202A) ALHNHYTQKS LSLSPGKGGG GSGGGGSGGGGSDSSPLLQF GGQVRQRYLY TDDACQTEAH LEIREDGTVG GAADQSPESL LQLKALKPGVIQILGVKTSR FLCQRPDGTL YGSLHFDPEA CSFRELLLED GYNVYQSEAH GLPLHLPCNRSPHRDPASRG RARFLPLPGL PPALPEPPGI LAPQPPDVGS SDPLAMVGGS QARSPSYAS *-Notethat the FGF21 wild-type sequence in this table refers to NCBI referencesequence number NP_061986.1 (SEQ ID NO: 1) unless otherwise specified.

All mutations in the FGF21 moiety and corresponding amino acid numberingof said mutations refers back to (SEQ ID NO:1) not to the full-lengthsequences in this table which may also include Fc and linker regions.

In specific aspects, provided herein are methods of treating, preventingor managing a metabolic disorder or cardiovascular disorder (e.g.,obesity, NALFD, NASH, or diabetes), said methods comprises administeringa human FGF21 protein variant, which is a fusion protein comprising ahuman Fc region (e.g., modified human Fc region) fused to a mature humanFGF21 protein or a fragment thereof comprising one or more mutationsselected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, andG202A, according to the number of SEQ ID NO: 1.

In specific aspects, provided herein are methods of treating, preventingor managing a metabolic disorder or cardiovascular disorder (e.g.,obesity, NALFD, NASH, or diabetes), said methods comprises administeringa human FGF21 protein variant, which is a fusion protein comprising ahuman Fc region (e.g., modified human Fc region) fused to a mature humanFGF21 protein or a fragment thereof, such as V103 comprising the aminoacid sequence of SEQ ID NO: 11.

Functional assays to determine or measure the bioactivity of FGF21 or avariant thereof or a fragment thereof have been described, for example,see PCT Publication No. WO 2013/049247, which is incorporated byreference herewith in its entirety. In specific aspects, functionalassays are in vitro assays. In other aspects, functional assays are invivo assays. In certain aspects, functional assays are ex vivo assays.

In specific aspects FGF21 downstream biomarkers can be assessed todetermine the bioactivity of FGF21 or a variant thereof or a fragmentthereof. A “FGF21 downstream biomarker,” as used herein, is a gene orgene product, or measurable indicia of a gene or gene product. In someaspects, a gene or activity that is a downstream marker of FGF21exhibits an altered level of expression, or in a vascular tissue. Insome aspects, an activity of the downstream marker is altered in thepresence of an FGF21 modulator. In some aspects, the downstream markersexhibit altered levels of expression when FGF21 is perturbed with anFGF21 modulator. FGF21 downstream markers include, without limitation,glucose or 2-deoxy-glucose uptake, pERK and other phosphorylated oracetylated proteins or NAD levels.

Pharmaceutical Compositions and Formulations

In specific aspects, provided herein are pharmaceutical compositionscomprising an FGF21 protein variants, for example FGF21 protein variantsdescribed in Table 1, e.g., V103 (SEQ ID NO: 11), for use in methods oftreating, preventing, and/or managing metabolic or cardiovasculardisorders, for example hypertriglyceridemia and cardiac risk, insulinresistance such as patients with genetic mutations of insulin receptorand lipodystrophy, diabetes, obesity, and nonalcoholic fatty liverdisease (NAFLD)/nonalcoholic steatohepatitis (NASH).

Pharmaceutical compositions described herein can comprise atherapeutically effective amount of an FGF21 protein variant and apharmaceutically or physiologically acceptable carrier. The carrier isgenerally selected to be suitable for the intended mode ofadministration and can include agents for modifying, maintaining, orpreserving, for example, the pH, osmolarity, viscosity, clarity, color,isotonicity, odor, sterility, stability, rate of dissolution or release,adsorption, and/or penetration of the composition. Typically, thesecarriers include aqueous or alcoholic/aqueous solutions, emulsions orsuspensions, including saline and/or buffered media.

Suitable agents for inclusion in the pharmaceutical compositionsinclude, but are not limited to, amino acids (such as glycine,glutamine, asparagine, arginine, histidine, or lysine), antimicrobials,antioxidants (such as ascorbic acid, sodium sulfite, or sodiumhydrogen-sulfite), buffers (such as borate, bicarbonate, Tris-HCl,citrates, phosphates, or other organic acids), bulking agents (such asmannitol or glycine), chelating agents (such as ethylenediaminetetraacetic acid (EDTA)), complexing agents (such as caffeine,polyvinylpyrrolidone, beta-cyclodextrin, orhydroxypropyl-beta-cyclodextrin), fillers, monosaccharides,disaccharides, and other carbohydrates (such as glucose, mannose, ordextrins), proteins (such as free serum albumin, gelatin, orimmunoglobulins), coloring, flavoring and diluting agents, emulsifyingagents, hydrophilic polymers (such as polyvinylpyrrolidone), lowmolecular weight polypeptides, salt-forming counterions (such assodium), preservatives (such as benzalkonium chloride, benzoic acid,salicylic acid, thimerosal, phenethyl alcohol, methylparaben,propylparaben, chlorhexidine, sorbic acid, or hydrogen peroxide),solvents (such as glycerin, propylene glycol, or polyethylene glycol),sugar alcohols (such as mannitol or sorbitol), suspending agents,surfactants or wetting agents (such as pluronics; PEG; sorbitan esters;polysorbates such as Polysorbate 20 or Polysorbate 80; Triton;tromethamine; lecithin; cholesterol or tyloxapal), stability enhancingagents (such as sucrose or sorbitol), tonicity enhancing agents (such asalkali metal halides, such as sodium or potassium chloride, or mannitolsorbitol), delivery vehicles, diluents, excipients and/or pharmaceuticaladjuvants

Parenteral vehicles include sodium chloride solution, Ringer's dextrose,dextrose and sodium chloride and lactated Ringer's. Suitablephysiologically-acceptable thickeners such as carboxymethylcellulose,polyvinylpyrrolidone, gelatin and alginates may be included. Intravenousvehicles include fluid and nutrient replenishers and electrolytereplenishers, such as those based on Ringer's dextrose. In some cases,it will be preferable to include agents to adjust tonicity of thecomposition, for example, sugars, polyalcohols such as mannitol,sorbitol, or sodium chloride in a pharmaceutical composition. Forexample, in many cases it is desirable that the composition issubstantially isotonic. Preservatives and other additives, such asantimicrobials, antioxidants, chelating agents and inert gases, may alsobe present. The precise formulation will depend on the route ofadministration. Additional relevant principle, methods and componentsfor pharmaceutical formulations are well known. (See, e.g., Allen, LoydV. Ed, (2012) Remington's Pharmaceutical Sciences, 22th Edition, whichis incorporated by reference for this purpose)

When parenteral administration is contemplated, the pharmaceuticalcompositions are usually in the form of a sterile, pyrogen-free,parenterally acceptable composition. A particularly suitable vehicle forparenteral injection is a sterile, isotonic solution, properlypreserved. The pharmaceutical composition can be in the form of alyophilizate, such as a lyophilized cake.

In certain aspects, a pharmaceutical composition provided herein (e.g.,a composition comprising FGF21 protein variant V103) is for subcutaneousadministration. Suitable formulation components and methods forsubcutaneous administration of polypeptide therapeutics (e.g.,antibodies, fusion proteins and the like) are known in the art. See,e.g., Published United States Patent Application No 2011/0044977 andU.S. Pat. Nos. 8,465,739 and 8,476,239, each of which is incorporated byreference for this purpose. Typically, pharmaceutical compositions forsubcutaneous administration contain suitable stabilizers (e.g., aminoacids, such as methionine, and or saccharides such as sucrose),buffering agents, and/or tonicifying agents.

In specific aspects, provided herein is a pharmaceutical compositioncomprising an FGF21 protein variant (for example, an FGF21 proteinvariant set forth in Table 1 such as V103) as a lyophilisate. Inparticular aspects, a pharmaceutical composition provided hereincomprising an FGF21 protein variant (for example, an FGF21 proteinvariant set forth in Table 1 such as V103) as a lyophilized formulationis reconstituted as a solution prior to administration (e.g.,subcutaneous administration) to a subject.

In particular aspects, a pharmaceutical composition provided hereincomprising an FGF21 protein variant (for example, an FGF21 proteinvariant set forth in Table 1 such as V103) is in the form of a solution(e.g., after reconstitution of a lyophilisate) comprising 100 mg/mL ofthe FGF21 protein variant such as V103. In some aspects, apharmaceutical composition provided herein comprising an FGF21 proteinvariant (for example, an FGF21 protein variant set forth in Table 1 suchas V103) is in the form of a solution (e.g., after reconstitution of alyophilisate) comprising at least about 50 mg/mL, 60 mg/mL, 70 mg/mL, 80mg/mL, 90 mg/mL, 100 mg/mL, 110 mg/mL, 120 mg/mL, 130 mg/mL, 140 mg/mL,or 150 mg/mL of the FGF21 protein variant such as V103. In some aspects,a pharmaceutical composition provided herein comprising an FGF21 proteinvariant (for example, an FGF21 protein variant set forth in Table 1 suchas V103) is in the form of a solution (e.g., after reconstitution of alyophilisate) comprising about 50 mg/mL to 150 mg/mL or 100 mg/mL to 150mg/mL of the FGF21 protein variant such as V103. In one aspect, apharmaceutical composition provided herein comprising an FGF21 proteinvariant (for example, an FGF21 protein variant set forth in Table 1 suchas V103) is in the form of a solution (e.g., after reconstitution of alyophilisate) comprising about 75 mg/mL to 150 mg/mL of V103, forexample 100 mg/mL V103 (SEQ ID NO: 11).

In some aspects, a pharmaceutical composition provided herein comprisingan FGF21 protein variant (for example, an FGF21 protein variant setforth in Table 1 such as V103) is in the form of a solution (e.g., afterreconstitution of a lyophilisate) comprising trometamol buffer, forexample, at a concentration of 10 mM, 20 mM, 30 mM, 40 mM, or 50 mMtrometamol buffer. In some aspects, a pharmaceutical compositionprovided herein comprising an FGF21 protein variant (for example, anFGF21 protein variant set forth in Table 1 such as V103) is in the formof a solution (e.g., after reconstitution of a lyophilisate) comprisingtrometamol buffer, for example, at a concentration of 10 mM to 50 mMtrometamol buffer, for example 30 mM trometamol buffer.

In some aspects, a pharmaceutical composition provided herein comprisingan FGF21 protein variant (for example, an FGF21 protein variant setforth in Table 1 such as V103) is in the form of a solution (e.g., afterreconstitution of a lyophilisate) comprising sucrose, for example, atleast about 200 mM, 210 mM, 220 mM, 230 mM, 240 mM, 250 mM, 260 mM, 270mM, 280 mM, 290 mM, or 300 mM sucrose. In some aspects, a pharmaceuticalcomposition provided herein comprising an FGF21 protein variant (forexample, an FGF21 protein variant set forth in Table 1 such as V103) isin the form of a solution (e.g., after reconstitution of a lyophilisate)comprising sucrose, for example, at least about 250 mM to 300 mMsucrose, for example 270 mM.

In some aspects, a pharmaceutical composition provided herein comprisingan FGF21 protein variant (for example, an FGF21 protein variant setforth in Table 1 such as V103) is in the form of a solution (e.g., afterreconstitution of a lyophilisate) comprising Polysorbate, such asPolysorbate 20. In specific aspects, a pharmaceutical compositionprovided herein comprising an FGF21 protein variant (for example, anFGF21 protein variant set forth in Table 1 such as V103) is in the formof a solution (e.g., after reconstitution of a lyophilisate) comprisingat least about 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%,0.09%, or 0.10% Polysorbate 20. In specific aspects, a pharmaceuticalcomposition provided herein comprising an FGF21 protein variant (forexample, an FGF21 protein variant set forth in Table 1 such as V103) isin the form of a solution (e.g., after reconstitution of a lyophilisate)comprising 0.02% to 0.10% Polysorbate 20, for example 0.06% Polysorbate20.

In some aspects, a pharmaceutical composition provided herein comprisingan FGF21 protein variant (for example, an FGF21 protein variant setforth in Table 1 such as V103) is in the form of a solution (e.g., afterreconstitution of a lyophilisate) at a pH in the range of 6.5 to 9, forexample a pH of 8.0. In some aspects, a pharmaceutical compositionprovided herein comprising an FGF21 protein variant (for example, anFGF21 protein variant set forth in Table 1 such as V103) is in the formof a solution (e.g., after reconstitution of a lyophilisate) at a pH ofat least about 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0 or 9.5.

In some aspects, a pharmaceutical composition provided herein comprisingan FGF21 protein variant (for example, an FGF21 protein variant setforth in Table 1 such as V103) is in the form of a solution (e.g., afterreconstitution of a lyophilisate) comprising trometamol buffer, sucrose,and Polysorbate 20.

In particular aspects, a pharmaceutical composition provided hereincomprising an FGF21 protein variant (for example, an FGF21 proteinvariant set forth in Table 1 such as V103) is in the form of a solution(e.g., after reconstitution of a lyophilisate) comprising about 80 mg/mLto 100 mg/mL of the FGF21 protein variant such as V103 (SEQ ID NO: 11),about 30 mM of trometamol buffer, 270 mM sucrose, and 0.06% Polysorbate20, at pH 7.5 to 8.5, for example pH 8.0.

In particular aspects, a pharmaceutical composition provided hereincomprising an FGF21 protein variant (for example, an FGF21 proteinvariant set forth in Table 1 such as V103) is in the form of a solution(e.g., after reconstitution of a lyophilisate) comprising about 100mg/mL of the FGF21 protein variant such as V103 (SEQ ID NO: 11), about30 mM of trometamol buffer, 270 mM sucrose, and 0.06% Polysorbate 20, atpH 8.0.

In certain aspects, provided herein is a method of preparing apharmaceutical composition comprising an FGF21 protein variant (forexample, an FGF21 protein variant set forth in Table 1 such as V103),said method comprises formulating the FGF21 protein variant as alyophilisate suitable for reconstitution into a solution.

In certain aspects, provided herein is a method of preparing apharmaceutical composition comprising an FGF21 protein variant (forexample, an FGF21 protein variant set forth in Table 1 such as V103),said method comprises reconstituting a lyophilisate into a solutiondescribed herein, for example, a solution comprising up to about 100mg/mL (e.g., about 50 mg/mL to about 100 mg/mL0 of the FGF21 proteinvariant such as V103 (SEQ ID NO: 11), about 30 mM of trometamol buffer,270 mM sucrose, and 0.06% Polysorbate 20, at pH 8.0.

In certain aspects, provided herein is a method of preparing apharmaceutical composition comprising an FGF21 protein variant (forexample, an FGF21 protein variant set forth in Table 1 such as V103),said method comprises preparing a solution comprising about 100 mg/mL ofthe FGF21 protein variant such as V103 (SEQ ID NO: 11), about 30 mM oftrometamol buffer, 270 mM sucrose, and 0.06% Polysorbate 20, at pH 8.0.

In a particular aspect, a method of preparing a pharmaceuticalcomposition described herein comprises the step of diluting a solutioncontaining an FGF21 protein variant (for example, an FGF21 proteinvariant set forth in Table 1 such as V103), to a concentration in therange of 10 mg/mL to 90 mg/mL. In a specific aspect, the solution isdiluted with saline solution, for example, 0.9% saline solution.

V. EXAMPLES

The following examples are intended to illustrate the invention and arenot to be construed as being limitations thereon. Abbreviations are usedas is conventional in the art.

Example 1

An Fc-FGF21 variant fusion protein, e.g., V103 comprising SEQ ID NO: 11,has been described, see, e.g., PCT Publication No. WO2013/049247. Forexample, V103 has been reported to induce 2-deoxyglucose update by mouse3T3L1 adipocytes, to induce ERK signaling in cell-based ERKphosphorylation assays, and to reduce total plasma glucose, plasmainsulin, and body weight in ob/ob mice (mice lacking functional leptinas a mouse model for type 2 diabetes). V103 has also been reported tohave a longer half-life and to be more thermodynamically stable, forexample as assessed by melting temperature.

The PK characteristics of V103 in cynomolgus monkeys have beendetermined in a 2-dose IV and SC injection toxicity study with a 4-weekrecovery period and in a 13-week IV and SC injection toxicity study witha 13-week recovery period, respectively.

Pharmacokinetics of V103 was similar in both studies, with no apparentgender differences. A moderate accumulation of V103 was observed in13-week GLP toxicity study (accumulation index 1.15-2.36 across the fourdose groups, as measured by Cmax and AUC 0-7 d after the first and lastdose).

Exposures to V103 (Cmax and AUC) increased approximatelydose-proportionally over the dose range of 0.3-100 mg/kg.Pharmacokinetics following intravenous was bi-exponential, with a linearterminal elimination phase. V103 was absorbed slowly after subcutaneousadministration (100 mg/kg) and the maximum exposure was observed 1-4days after the SC administration. The terminal elimination phase was notabsorption limited and estimated subcutaneous bioavailability was60-90%. Table 2 below summarizes the mean toxicokinetic parameterscalculated for the penultimate (Day 85) and the ultimate dose (Day 92)of the 13-week GLP-compliant toxicology study.

TABLE 2 Mean TK parameters obtained for the penultimate (Day 85) and theultimate dose (Day 92) of the 13-week GLP-compliant toxicity study incynomolgus monkey Group 2 Group 3 Group 4 Group 5 Study TK 0.3 mg/kg iv3.0 mg/kg iv 30 mg/kg iv 30 mg/kg sc day parameter Mean ± SD Mean ± SDMean ± SD Mean ± SD 85 Tmax (h) 0.25 0.00 5.50 9.58 1.54 3.16 22.7 12.685 Cmax (μg/mL) 20.7 4.20 231 87.0 2290 1110 1440 257 85 Cmax/Dose 69.014.0 76.9 28.9 76.4 37.0 48.1 8.55 85 AUC0-7 d 1990 408 19200 6680189000 96100 186000 31200 85 AUC0-7 d/Dose 6630 1360 6390 2230 6300 31906190 1040 92 Tmax (h) 2.19 3.88 48.0 27.7 92 Cmax (μg/mL) 2020 256 1190158 92 Cmax/Dose 67.3 8.52 39.5 5.25 92 AUC0-7 d 189000 6920 15400033300 92 AUC0-7 d/Dose 6310 231 5140 1110 Day 85, n = 6 animals per maingroup 0.3, 3.0, 30 mg/kg iv and 30 mg/kg sc V103. Day 92, n = 4 animalsper recovery group 30 mg/kg iv and 30 mg/kg sc V103. Cmax/Dose((μg/mL)/(mg/kg)); AUC0-7 d (μg × h/mL); AUC0-7 d/Dose ((μg ×h/mL)/(mg/kg)) The GLP validated MSD-based PK assay used fordetermination of V103 concentrations in cynomolgus monkey serumquantifies the bioactive but partially also the truncated/inactive formsof V103.

In agreement with the rat pharmacokinetics, HPLC-MS/MS assessmentsconfirmed that in vivo in cynomolgus monkeys, the central part of FGF21subdomain of V103 is more stable (T_(1/2)˜9-12 days) than the C-terminalsubdomain, required for FGF21 receptor binding and activity (T_(1/2)˜5-8days). Table 3 below summarizes the pharmacokinetic properties ofbioactive and truncated/inactive forms of V103 determined in the 13-weekGLP-compliant toxicology study.

TABLE 3 In-vivo integrity of V103 following weekly IV and SCadministration in the 13-week GLP-compliant toxicity study in cynomolgusmonkeys V103 (C-terminal V103 (Central TK parameter V103 ^(a) peptide ofFGF21) ^(b) peptide of FGF21) ^(b) Group 5 AUC* 423000 224000 483000 30mg/kg SC T½ (Days) 10.0 8.2 12.3 TK parameters were calculated from themean TK-profiles after the last dose (Day 92) *AUC0-90 days (μg × h/mL)^(a) as measured by the MSD-based assay ^(b) as measured by theLC-MS/MS-based assay

Anti-V103 antibodies (ADAs) were observed in a number of animals. Inmost cases, the treatment-related ADA response was associated with anaccelerated clearance of V103, however, the overall impact on total andsustained exposure to V103 in these studies was minimal. Selectivity ofADA response was assessed during the 13-week GLP toxicity study and noneof the ADA positive samples was cross-reactive with the endogenouswild-type FGF21.

Example 2

The clinical study purpose is to evaluate the safety and tolerability ofmultiple doses of V103 administered subcutaneously over 3 months inobese subjects. In addition, the study will also determine earlyefficacy signals in various metabolic diseases associated with elevatedtriglycerides and/or obesity, and whether V103 displays the clinicalsafety and efficacy profile as a therapeutic suitable for development inthe metabolic diseases of hypertriglyceridemia, obesity, and/ornon-alcoholic fatty liver disease.

Objectives and Endpoints

The primary objective(s) is to assess the safety and tolerability inobese subjects following repeated dosing of V103 by subcutaneous (SC)injection over 12 weeks. Endpoints related to this objective include:

-   -   Adverse events (AE)    -   Vital signs    -   Liver function tests aspartate transaminase (AST), alanine        transaminase (ALT), total bilirubin (Total Bili) and alkaline        phosphatase (ALP)    -   24 hour urinary cortisol    -   Electrolytes

Secondary objectives include:

-   -   To assess the effects of V103 on triglycerides and lipid        profiles at 12 weeks of treatment.    -   To assess the potential effects of V103 on bone biomarkers of        resorption, bone formation and balance of resorption to        deposition at 12 weeks of treatment.    -   To assess the effects of V103 on weight as measured by body        weight, body mass index (BMI) and waist circumference at 12        weeks.

Endpoints related to these objectives include:

-   -   Total cholesterol, LDL-C, HDL-C and Triglyceride (fasting) at 12        weeks    -   Biomarkers of bone resorption (serum CTX-1, urine NTX-1) at 12        weeks    -   and Biomarkers bone formation (serum BSAP, P1NP, and        osteocalcin) at 12 weeks    -   Weight, Body mass index (BMI), waist circumference, and percent        liver fat fraction as measured by MRI at 12 weeks

Other study objectives include:

-   -   To assess pharmacokinetics (PK) of V103 in obese subjects        following repeated dosing over 12 weeks.    -   To assess immunogenicity of V103 after repeat SC dosing of V103.    -   To assess the potential glycemic metabolic effects of V103.

Non-limiting examples of endpoints related to these other studyobjectives include:

-   -   PK parameters will be determined, including: AUClast, Cmax, and        Tmax;    -   Fasting glucose, insulin, glucagon, and C-peptide, and derived        estimates of insulin sensitivity and secretion (HOMA) up to 12        weeks;    -   Hb1c, glycated albumin;    -   Adiponectin; and    -   Pre and post-dose anti-drug antibodies (ADAs) over 26 weeks.

Study Design:

The study is designed as a non-confirmatory, multicenter, randomized,investigator- and subject-blinded, placebo-controlled safety study inobese subjects with V103 or placebo administered subcutaneously withrepeated dosing over 3 months.

Approximately 60 subjects will be randomized (1:1, active:placebo) toreceive V103 300 mg q 4 weeks for three doses by subcutaneous (SC)injection, or a matching placebo.

The trial consists of three main periods: (1) screening, which will lastranging from 1 to 4 weeks, (2) a subject- and investigator-blindedrandomized treatment period, projected to last 12 weeks, and (3) anend-of-study evaluation, about 120 days (approximately 6 half-lives)after last dose of study drug.

Cohort expansion: After interim analysis (e.g., after approximately 15subjects in each group reached the end of treatment (Day 84)), a lowerdose cohort may be evaluated if there is evidence of efficacy and a needto evaluate additional doses to inform future studies prior topopulation expansion.

Population:

The study population will be comprised of approximately 60 adult maleand female obese subjects 18 to 55 years of age (inclusive).

Key Inclusion criteria:

-   -   Male and female subjects 18 to 55 years of age inclusive;    -   Body mass index (BMI) within the range of 30 to 45 kg/m²,        inclusive, with ethnic adjustment ≥27.5 for Asian individuals        (WHO Expert Consultation, 2004, Lancet, 363(9403):157-63);        BMI=Body weight (kg)/[Height (m)]², inclusive; and    -   Triglyceride 150-500 mg/dL (1.69-5.65 mmol/L), inclusive, at        screening.

Key Exclusion criteria:

-   -   History of hepatobilliary disease, cholelithiasis, or biliary        sludge by history or at screening ultrasound, hepatic        encephalopathy, esophageal varices, or porticaval shunt;    -   Liver disease or liver injury as indicated by abnormal liver        function tests (ALT, AST, GGT, alkaline phosphatase, or serum        bilirubin) above upper limit of normal at screening or baseline;    -   Chronic infection with Human Immunodeficiency Virus (HIV),        Hepatitis B (HBV) or Hepatitis C (HCV). A positive HBV surface        antigen (HBsAg) test, or if standard local practice, a positive        HBV core antigen test, excludes a subject. Subjects with a        positive HCV antibody test should have HCV RNA levels measured.        Subjects with positive (detectable) HCV RNA should be excluded;    -   Fasting triglycerides greater than or equal to 500 mg/dL [5.65        mmol/L], or concomitant use of drug treatment for        hypertriglyceridemia (fibrates, omega-3 fatty acids, nicotinic        acid);    -   History of pancreatic injury or pancreatitis, or other        pancreatic disease. Amylase or lipase above ULN at screening or        baseline;    -   History of hypersensitivity to drugs of similar biological        class, FGF21 protein analogue, or Fc fusion proteins;    -   History of bone disorders including but not limited to        osteoporosis, osteopenia, osteomalacia, severe vitamin D        deficiency;    -   Plasma 25-hydroxyvitamin D level is below the lower limit of the        normal range at screening;    -   Contraindications to MRI;    -   Change in body weight (more than 5% self-reported OR 5 kg        self-reported change during the previous 3 months);    -   Use of weight loss drugs: Orlistat (Xenical, Alli), lorcaserin        (Belviq), phentermine-topiramate (Qsymia), naltrexone-bupropion        (Contrave), or liraglutide (Victoza or Saxenda) or other        glucagon-like peptide-1 (GLP1) receptor agonists (exenatide        (Byetta/Bydureon), lixisenatide (Luxumia), albiglutide (Tanzeum)        or dulaglutide (Trulicity) or others); and    -   Enrollment in a diet, weight loss or exercise programs with the        specific intent of losing weight, within 3 months prior to        randomization, OR clinical diagnosis of any eating disorder is        also an exclusion.

Pharmacokinetic assessments: Cmax, Tmax, Clast, Tlast, AUClast, AUCtau,AUCinf, T1/2, Vz/F and CL/F from the serum concentration-time data, iffeasible

Efficacy/PD assessments:

-   -   Fasting triglycerides    -   Body weight    -   Key safety assessments    -   Adverse event and Serious adverse event monitoring    -   Bone biomarkers    -   Liver function    -   Ultrasound    -   Pancreatic function    -   Pituitary-adrenal function    -   Other pituitary-endocrine safety monitoring    -   Physical examinations and vital signs    -   Monitoring of routine laboratory markers in blood and urine    -   Electrocardiogram    -   Columbia-Suicide Severity Rating Scale (C-SSRS)    -   Mechanistic biomarkers and biomarkers of inflammation

Additional Embodiments

In one aspect, provided herein is a method of treating, preventing, ormanaging a metabolic disorder or a cardiovascular disorder, comprisingadministering to a subject in need thereof a human FGF21 protein variantat a dose in the range of 100 mg to 600 mg.

In another aspect, provided herein is a method of treating, preventing,or managing hypercholesterolemia, mixed dyslipidemia, orhypertriglyceridemia, comprising administering to a subject in needthereof a human FGF21 protein variant at a dose in the range of 100 mgto 600 mg.

In another aspect, provided herein is a method of reducing body weight,comprising administering to a subject in need thereof of a human FGF21protein variant at a dose in the range of 100 mg to 600 mg.

In another aspect, provided herein is a method of reducing elevatedLDL-C, total-C, triglyceride, and/or Apo B, comprising administering toa subject in need thereof a human FGF21 protein variant at a dose in therange of 100 mg to 600 mg.

In another aspect, provided herein is a method of increasing HDL-C,comprising administering to a subject in need thereof a human FGF21protein variant at a dose in the range of 100 mg to 600 mg.

In another aspect, provided herein is a method of reducing triglyceridelevels, comprising administering to a subject in need thereof a humanFGF21 protein variant at a dose in the range of 100 mg to 600 mg.

In another aspect, provided herein is a method of reducingcardiovascular risk, comprising administering to a subject in needthereof a human FGF21 protein variant at a dose in the range of 100 mgto 600 mg.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing a metabolic disorder ora cardiovascular disorder, said method comprising administering to asubject in need thereof the human FGF21 protein variant at a dose in therange of 100 mg to 600 mg.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing hypercholesterolemia,mixed dyslipidemia, or hypertriglyceridemia, said method comprisingadministering to a subject in need thereof the human FGF21 proteinvariant at a dose in the range of 100 mg to 600 mg.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of reducing body weight, said method comprisingadministering to a subject in need thereof of the human FGF21 proteinvariant at a dose in the range of 100 mg to 600 mg.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of reducing elevated LDL-C, total-C, triglyceride, and/orApo B, comprising administering to a subject in need thereof the humanFGF21 protein variant at a dose in the range of 100 mg to 600 mg.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of increasing HDL-C, said method comprising administering toa subject in need thereof the human FGF21 protein variant at a dose inthe range of 100 mg to 600 mg.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of reducing triglyceride levels, said method comprisingadministering to a subject in need thereof the human FGF21 proteinvariant at a dose in the range of 100 mg to 600 mg.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of reducing cardiovascular risk, said method comprisingadministering to a subject in need thereof the human FGF21 proteinvariant at a dose in the range of 100 mg to 600 mg.

In one aspect, provided herein is a use of a human FGF21 protein variantin the preparation of a medicament for use in a method of treating,preventing, or managing a metabolic disorder or a cardiovasculardisorder, said method comprising administering to a subject in needthereof the human FGF21 protein variant at a dose in the range of 100 mgto 600 mg.

In one aspect, provided herein is a use of a human FGF21 protein variantin the preparation of a medicament for use in a method of treating,preventing, or managing hypercholesterolemia, mixed dyslipidemia, orhypertriglyceridemia, said method comprising administering to a subjectin need thereof the human FGF21 protein variant at a dose in the rangeof 100 mg to 600 mg.

In one aspect, provided herein is a use of a human FGF21 protein variantin the preparation of a medicament for use in a method of reducing bodyweight, said method comprising administering to a subject in needthereof of the human FGF21 protein variant at a dose in the range of 100mg to 600 mg.

In one aspect, provided herein is a use of a human FGF21 protein variantin the preparation of a medicament for use in a method of reducingelevated LDL-C, total-C, triglyceride, and/or Apo B, comprisingadministering to a subject in need thereof the human FGF21 proteinvariant at a dose in the range of 100 mg to 600 mg.

In one aspect, provided herein is a use of a human FGF21 protein variantin the preparation of a medicament for use in a method of increasingHDL-C, said method comprising administering to a subject in need thereofthe human FGF21 protein variant at a dose in the range of 100 mg to 600mg.

In one aspect, provided herein is a use of a human FGF21 protein variantin the preparation of a medicament for use in a method of reducingtriglyceride levels, said method comprising administering to a subjectin need thereof the human FGF21 protein variant at a dose in the rangeof 100 mg to 600 mg.

In one aspect, provided herein is a use of a human FGF21 protein variantin the preparation of a medicament for use in a method of reducingcardiovascular risk, said method comprising administering to a subjectin need thereof the human FGF21 protein variant at a dose in the rangeof 100 mg to 600 mg.

In one aspect, provided herein is a use of a human FGF21 protein variantin the preparation of a medicament for use in a method of reducingcardiovascular risk, said method comprising administering to a subjectin need thereof the human FGF21 protein variant at a dose in the rangeof 100 mg to 600 mg.

In one aspect, provided herein is a use of a human FGF21 protein variantat an amount in the range of 100 mg to 600 mg in the preparation of amedicament for use in a method of treating, preventing, or managing ametabolic disorder or a cardiovascular disorder.

In one aspect, provided herein is a use of a human FGF21 protein variantat an amount in the range of 100 mg to 600 mg in the preparation of amedicament for use in a method of treating, preventing, or managinghypercholesterolemia, mixed dyslipidemia, or hypertriglyceridemia.

In one aspect, provided herein is a use of a human FGF21 protein variantat an amount in the range of 100 mg to 600 mg in the preparation of amedicament for use in a method of reducing body weight.

In one aspect, provided herein is a use of a human FGF21 protein variantat an amount in the range of 100 mg to 600 mg in the preparation of amedicament for use in a method of reducing elevated LDL-C, total-C,triglyceride, and/or Apo B.

In one aspect, provided herein is a use of a human FGF21 protein variantat an amount in the range of 100 mg to 600 mg in the preparation of amedicament for use in a method of increasing HDL-C.

In one aspect, provided herein is a use of a human FGF21 protein variantat an amount in the range of 100 mg to 600 mg in the preparation of amedicament for use in a method of reducing triglyceride levels.

In one embodiment, the method reduces triglyceride levels by at leastabout 40% or at least about 50%.

In one aspect, provided herein is a use of a human FGF21 protein variantat an amount in the range of 100 mg to 600 mg in the preparation of amedicament for use in a method of reducing cardiovascular risk.

In one aspect, provided herein is a use of a human FGF21 protein variantat an amount in the range of 100 mg to 600 mg in the manufacture of amedicament for use in treating, preventing, or managing a metabolicdisorder or a cardiovascular disorder.

In one aspect, provided herein is a use of a human FGF21 protein variantat an amount in the range of 100 mg to 600 mg in the manufacture of amedicament for use in treating, preventing, or managinghypercholesterolemia, mixed dyslipidemia, or hypertriglyceridemia.

In one aspect, provided herein is a use of a human FGF21 protein variantat an amount in the range of 100 mg to 600 mg in the manufacture of amedicament for use in reducing body weight.

In one aspect, provided herein is a use of a human FGF21 protein variantat an amount in the range of 100 mg to 600 mg in the manufacture of amedicament for use in reducing elevated LDL-C, total-C, triglyceride,and/or Apo B.

In one aspect, provided herein is a use of a human FGF21 protein variantat an amount in the range of 100 mg to 600 mg in the manufacture of amedicament for use in increasing HDL-C.

In one aspect, provided herein is a use of a human FGF21 protein variantat an amount in the range of 100 mg to 600 mg in the manufacture of amedicament for use in reducing triglyceride levels.

In one embodiment, the method reduces triglyceride levels by at leastabout 40% or at least about 50%.

In one aspect, provided herein is a use of a human FGF21 protein variantat an amount in the range of 100 mg to 600 mg in the manufacture of amedicament for use in reducing cardiovascular risk.

In one aspect, provided herein is a use of a human FGF21 protein variantin the manufacture of a medicament for the treatment of a metabolicdisorder or a cardiovascular disorder, wherein the unit dose of thehuman FGF21 protein variant is in the range of 100 mg to 600 mg.

In one aspect, provided herein is a use of a human FGF21 protein variantin the manufacture of a medicament for the treatment ofhypercholesterolemia, mixed dyslipidemia, or hypertriglyceridemia,wherein the unit dose of the human FGF21 protein variant is in the rangeof 100 mg to 600 mg.

In one aspect, provided herein is a use of a human FGF21 protein variantin the manufacture of a medicament for use in reducing body weight,wherein the unit dose of the human FGF21 protein variant is in the rangeof 100 mg to 600 mg.

In one aspect, provided herein is a use of a human FGF21 protein variantin the manufacture of a medicament for use in reducing LDL-C, total-C,triglyceride, and/or Apo B levels, wherein the unit dose of the humanFGF21 protein variant is in the range of 100 mg to 600 mg.

In one aspect, provided herein is a use of a human FGF21 protein variantin the manufacture of a medicament for use in a method of increasingHDL-C, wherein the unit dose of the human FGF21 protein variant is inthe range of 100 mg to 600 mg.

In one aspect, provided herein is a use of a human FGF21 protein variantin the manufacture of a medicament for use in reducing triglyceridelevels, wherein the unit dose of the human FGF21 protein variant is inthe range of 100 mg to 600 mg.

In one aspect, provided herein is a use of a human FGF21 protein variantin the manufacture of a medicament for use in a method of reducingcardiovascular risk, wherein the unit dose of the human FGF21 proteinvariant is in the range of 100 mg to 600 mg.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing a metabolic disorder ora cardiovascular disorder in a human subject, wherein the human FGF21protein variant is provided for administration at a dose in the range of100 mg to 600 mg. In one aspect, provided herein is a human FGF21protein variant for use in a method of treating, preventing, or managinga metabolic disorder or a cardiovascular disorder in a human subject,wherein the human FGF21 protein variant is provided for administrationat an amount in the range of 100 mg to 600 mg.

In some embodiments of these aspects, the metabolic disorder orcardiovascular disorder is selected from hypercholesterolemia,dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease(NAFLD), nonalcoholic steatohepatitis (NASH), type 2 diabetes, andobesity.

In some embodiments of these aspects, treating, preventing, or managingthe metabolic disorder or cardiovascular disorder comprises or ischaracterized by reducing one or more of the following: body weight,liver fat content, elevated LDL-C, total-C, triglyceride, and Apo Blevels in the subject. In certain embodiments, treating, preventing, ormanaging the metabolic disorder or cardiovascular disorder comprises oris characterized by increasing HDL-C levels in the subject.

In some embodiments of these aspects, treating, preventing, or managingthe metabolic disorder or cardiovascular disorder comprises or ischaracterized by reducing triglyceride levels in the subject by at leastabout 40% or at least about 50%. In some embodiments, treating,preventing, or managing the metabolic disorder or cardiovasculardisorder comprises or is characterized by reducing triglyceride levelsin the subject by at least about 40%. In some embodiments, treating,preventing, or managing the metabolic disorder or cardiovasculardisorder comprises or is characterized by reducing triglyceride levelsin the subject by at least about 50%.

In some embodiments of these aspects, treating, preventing, or managingthe metabolic disorder or cardiovascular disorder comprises or ischaracterized by reducing cardiovascular risk in the subject.

In some embodiments of these aspects, the metabolic disorder orcardiovascular disorder is dyslipidemia, optionally mixed dyslipidemia,hypertriglyceridemia, optionally severe hypertriglyceridemia, orhypercholesterolemia, optionally primary hypercholesterolemia. In someembodiments, the metabolic disorder or cardiovascular disorder isnonalcoholic fatty liver disease (NAFLD) or nonalcoholic steatohepatitis(NASH). In certain embodiments, the subject is 18 to 55 years of age. Insome embodiments, the subject has a body mass index (BMI) within therange of 30 to 45 kg/m², inclusive, with ethnic adjustment ≥27.5 for asubject of Asian descent or Asian ancestry. In some embodiments, thesubject has a body mass index (BMI) within the range of 30 to 45 kg/m²,inclusive, optionally wherein the BMI is ≥27.5 for a subject of Asiandescent or Asian ancestry. In some embodiments, the subject hastriglyceride levels in the range of 150-500 mg/dL (1.69-5.65 mmol/L)when measured prior to administration of the human FGF21 proteinvariant.

In some embodiments of these aspects, the human FGF21 variant is afusion protein comprising a human Fc region fused to a mature humanFGF21 protein or a fragment thereof comprising one or more mutationsselected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202Aaccording to the numbering of SEQ ID NO: 1. In certain embodiments, thehuman FGF21 protein variant comprises the amino acid sequence of SEQ IDNO: 11.

In some embodiments of these aspects, the human FGF21 protein variant isprovided for administration at a dose of at least 100 mg, 150 mg, 200mg, 250 mg, 300 mg, 350 mg, or 400 mg. In some embodiments, the humanFGF21 protein variant is provided for administration at a dose ofapproximately 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg,170 mg, 180 mg, or 190 mg, optionally wherein the human FGF21 proteinvariant is provided at a dose of approximately 100 mg or 150 mg. In someembodiments, the human FGF21 protein variant is provided foradministration at a dose of approximately 200 mg, 210 mg, 220 mg, 230mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320mg, 330 mg, 340 mg, or 350 mg, optionally wherein the human FGF21protein variant is provided at a dose of approximately 200, 250 mg, or300 mg. In some embodiments, the human FGF21 protein variant is providedfor administration at a dose of at least 100 mg. In some embodiments,the human FGF21 protein variant is provided for administration at a doseof approximately 100 mg. In some embodiments, the human FGF21 proteinvariant is provided for administration at a dose of at least 150 mg. Insome embodiments, the human FGF21 protein variant is provided foradministration at a dose of approximately 150 mg. In some embodiments,the human FGF21 protein variant is provided for administration at a doseof at least 200 mg. In some embodiments, the human FGF21 protein variantis provided for administration at a dose of approximately 200 mg. Insome embodiments, the human FGF21 protein variant is provided foradministration at a dose of at least 250 mg. In some embodiments, thehuman FGF21 protein variant is provided for administration at a dose ofapproximately 250 mg. In some embodiments, the human FGF21 proteinvariant is provided for administration at a dose of 250 mg. In someembodiments, the human FGF21 protein variant is provided foradministration at a dose of at least 300 mg. In some embodiments, thehuman FGF21 protein variant is provided for administration at a dose ofapproximately 300 mg. In some embodiments, the human FGF21 proteinvariant is provided for administration at a dose of 300 mg.

In some embodiments of these aspects, the human FGF21 protein variant isprovided for administration at an amount of at least 100 mg, 150 mg, 200mg, 250 mg, 300 mg, 350 mg, or 400 mg. In some embodiments, the humanFGF21 protein variant is provided for administration at an amount ofapproximately 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg,170 mg, 180 mg, or 190 mg, optionally wherein the human FGF21 proteinvariant is provided at a amount of approximately 100 mg or 150 mg. Insome embodiments, the human FGF21 protein variant is provided foradministration at an amount of approximately 200 mg, 210 mg, 220 mg, 230mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320mg, 330 mg, 340 mg, or 350 mg, optionally wherein the human FGF21protein variant is provided at an amount of approximately 200 mg, 250mg, or 300 mg. In some embodiments, the human FGF21 protein variant isprovided for administration at an amount of at least 100 mg. In someembodiments, the human FGF21 protein variant is provided foradministration at an amount of approximately 100 mg. In someembodiments, the human FGF21 protein variant is provided foradministration at an amount of at least 150 mg. In some embodiments, thehuman FGF21 protein variant is provided for administration at an amountof approximately 150 mg. In some embodiments, the human FGF21 proteinvariant is provided for administration at an amount of at least 200 mg.In some embodiments, the human FGF21 protein variant is provided foradministration at an amount of approximately 200 mg. In someembodiments, the human FGF21 protein variant is provided foradministration at an amount of at least 250 mg. In some embodiments, thehuman FGF21 protein variant is provided for administration at an amountof approximately 250 mg. In some embodiments, the human FGF21 proteinvariant is provided for administration at an amount of 250 mg. In someembodiments, the human FGF21 protein variant is provided foradministration at an amount of at least 300 mg. In some embodiments, thehuman FGF21 protein variant is provided for administration at an amountof approximately 300 mg. In some embodiments, the human FGF21 proteinvariant is provided for administration at an amount of 300 mg.

In some embodiments, the human FGF21 protein variant is provided in aform for subcutaneous administration. In some embodiments, the humanFGF21 protein variant is provided for administration once a month oronce every 4 weeks, once every 3 weeks, once every 2 weeks, or onceevery week. In some embodiments, the human FGF21 protein variant isprovided in a form for administration once a month or once every 4weeks, once every 3 weeks, once every 2 weeks, or once every week. Insome embodiments, the human FGF21 protein variant is provided in a formto be administered subcutaneously.

In one aspect, provided herein is a method of treating, preventing, ormanaging a metabolic disorder or a cardiovascular disorder in a humansubject, wherein the method comprises administering a human FGF21protein variant at a dose in the range of 100 mg to 600 mg to thesubject. In one aspect, provided herein is a method of treating,preventing, or managing a metabolic disorder or a cardiovasculardisorder in a human subject, wherein the method comprises administeringa human FGF21 protein variant at an amount in the range of 100 mg to 600mg to the subject.

In some embodiments, the metabolic disorder or cardiovascular disorderis selected from hypercholesterolemia, dyslipidemia,hypertriglyceridemia, nonalcoholic fatty liver disease (NAFLD),nonalcoholic steatohepatitis (NASH), type 2 diabetes, and obesity.

In some embodiments, treating, preventing, or managing the metabolicdisorder or cardiovascular disorder comprises or is characterized byreducing one or more of the following: body weight, liver fat content,elevated LDL-C, total-C, triglyceride, and Apo B levels in the subject.In certain embodiments, treating, preventing, or managing the metabolicdisorder or cardiovascular disorder comprises or is characterized byincreasing HDL-C levels in the subject.

In some embodiments, treating, preventing, or managing the metabolicdisorder or cardiovascular disorder comprises or is characterized byreducing triglyceride levels in the subject by at least about 40% or atleast about 50%. In some embodiments, treating, preventing, or managingthe metabolic disorder or cardiovascular disorder comprises or ischaracterized by reducing triglyceride levels in the subject by at leastabout 40%. In some embodiments, treating, preventing, or managing themetabolic disorder or cardiovascular disorder comprises or ischaracterized by reducing triglyceride levels in the subject by at leastabout 50%.

In some embodiments, treating, preventing, or managing the metabolicdisorder or cardiovascular disorder comprises or is characterized byreducing cardiovascular risk in the subject.

In some embodiments, the metabolic disorder or cardiovascular disorderis dyslipidemia, hypertriglyceridemia, or hypercholesterolemia. In someembodiments, the metabolic disorder or cardiovascular disorder isdyslipidemia. In some embodiments, the metabolic disorder orcardiovascular disorder is hypertriglyceridemia. In some embodiments,the metabolic disorder or cardiovascular disorder ishypercholesterolemia. In some embodiments, the metabolic disorder orcardiovascular disorder is mixed dyslipidemia. In some embodiments, themetabolic disorder or cardiovascular disorder is severehypertriglyceridemia. In some embodiments, the metabolic disorder orcardiovascular disorder is primary hypercholesterolemia. In someembodiments, the metabolic disorder or cardiovascular disorder isnonalcoholic fatty liver disease (NAFLD) or nonalcoholic steatohepatitis(NASH). In some embodiments, the metabolic disorder or cardiovasculardisorder is nonalcoholic fatty liver disease (NAFLD). In someembodiments, the metabolic disorder or cardiovascular disorder isnonalcoholic steatohepatitis (NASH). In certain embodiments, the subjectis 18 to 55 years of age. In some embodiments, the subject has a bodymass index (BMI) within the range of 30 to 45 kg/m², inclusive, withethnic adjustment ≥27.5 for a subject of Asian descent or Asianancestry. In some embodiments, the subject has a body mass index (BMI)

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing hypercholesterolemia ina human subject, wherein the human FGF21 protein variant is provided foradministration at a dose or an amount in the range of 100 mg, 110 mg,120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg,210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg,300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, optionally whereinthe human FGF21 protein variant is provided at a dose of approximately100 mg, 150 mg, 200 mg, 250 mg, or 300 mg, and wherein the human FGF21variant is a fusion protein comprising a human Fc region fused to amature human FGF21 protein or a fragment thereof comprising one or moremutations selected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G,and G202A according to the numbering of SEQ ID NO: 1, optionally whereinthe human FGF21 protein variant comprises the amino acid sequence of SEQID NO: 11. In one embodiment thereof, the human FGF21 variant isprovided at a dose of approximately 100 mg, 150 mg, 200 mg, 250 mg, or300 mg for administration once per week. In one embodiment thereof, thehuman FGF21 variant is provided at a dose of approximately 100 mg, 150mg, 200 mg, 250 mg, or 300 mg for administration once every two weeks.In one embodiment thereof, the human FGF21 variant is provided at a doseof approximately 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg foradministration once every three weeks. In one embodiment thereof, thehuman FGF21 variant is provided at a dose of approximately 100 mg, 150mg, 200 mg, 250 mg, or 300 mg for administration once every four weeksor once per month.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing dyslipidemia in a humansubject, wherein the human FGF21 protein variant is provided foradministration at a dose or an amount in the range of 100 mg, 110 mg,120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg,210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg,300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, optionally whereinthe human FGF21 protein variant is provided at a dose of approximately100 mg, 150 mg, 200 mg, 250 mg, or 300 mg, and wherein the human FGF21variant is a fusion protein comprising a human Fc region fused to amature human FGF21 protein or a fragment thereof comprising one or moremutations selected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G,and G202A according to the numbering of SEQ ID NO: 1, optionally whereinthe human FGF21 protein variant comprises the amino acid sequence of SEQID NO: 11. In one embodiment thereof, the human FGF21 variant isprovided at a dose of approximately 100 mg, 150 mg, 200 mg, 250 mg, or300 mg for administration once per week. In one embodiment thereof, thehuman FGF21 variant is provided at a dose of approximately 100 mg, 150mg, 200 mg, 250 mg, or 300 mg for administration once every two weeks.In one embodiment thereof, the human FGF21 variant is provided at a doseof approximately 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg foradministration once every three weeks. In one embodiment thereof, thehuman FGF21 variant is provided at a dose of approximately 100 mg, 150mg, 200 mg, 250 mg, or 300 mg for administration once every four weeksor once per month.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing hypertriglyceridemia ina human subject, wherein the human FGF21 protein variant is provided foradministration at a dose or an amount in the range of 100 mg, 110 mg,120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg,210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg,300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, optionally whereinthe human FGF21 protein variant is provided at a dose of approximately100 mg, 150 mg, 200 mg, 250 mg, or 300 mg, and wherein the human FGF21variant is a fusion protein comprising a human Fc region fused to amature human FGF21 protein or a fragment thereof comprising one or moremutations selected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G,and G202A according to the numbering of SEQ ID NO: 1, optionally whereinthe human FGF21 protein variant comprises the amino acid sequence of SEQID NO: 11. In one embodiment thereof, the human FGF21 variant isprovided at a dose of approximately 100 mg, 150 mg, 200 mg, 250 mg, or300 mg for administration once per week. In one embodiment thereof, thehuman FGF21 variant is provided at a dose of approximately 100 mg, 150mg, 200 mg, 250 mg, or 300 mg for administration once every two weeks.In one embodiment thereof, the human FGF21 variant is provided at a doseof approximately 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg foradministration once every three weeks. In one embodiment thereof, thehuman FGF21 variant is provided at a dose of approximately 100 mg, 150mg, 200 mg, 250 mg, or 300 mg for administration once every four weeksor once per month.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) in a human subject, wherein the human FGF21protein variant is provided for administration at a dose or an amount inthe range of 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or350 mg, optionally wherein the human FGF21 protein variant is providedat a dose of approximately 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg,and wherein the human FGF21 variant is a fusion protein comprising ahuman Fc region fused to a mature human FGF21 protein or a fragmentthereof comprising one or more mutations selected from: Q55C, R105K,G148C, K150R, P158S, S195A, P199G, and G202A according to the numberingof SEQ ID NO: 1, optionally wherein the human FGF21 protein variantcomprises the amino acid sequence of SEQ ID NO: 11. In one embodimentthereof, the human FGF21 variant is provided at a dose of approximately100 mg, 150 mg, 200 mg, 250 mg, or 300 mg for administration once perweek. In one embodiment thereof, the human FGF21 variant is provided ata dose of approximately 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg foradministration once every two weeks. In one embodiment thereof, thehuman FGF21 variant is provided at a dose of approximately 100 mg, 150mg, 200 mg, 250 mg, or 300 mg for administration once every three weeks.In one embodiment thereof, the human FGF21 variant is provided at a doseof approximately 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg foradministration once every four weeks or once per month.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholicsteatohepatitis (NASH) in a human subject, wherein the human FGF21protein variant is provided for administration at a dose or an amount inthe range of 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or350 mg, optionally wherein the human FGF21 protein variant is providedat a dose of approximately 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg,and wherein the human FGF21 variant is a fusion protein comprising ahuman Fc region fused to a mature human FGF21 protein or a fragmentthereof comprising one or more mutations selected from: Q55C, R105K,G148C, K150R, P158S, S195A, P199G, and G202A according to the numberingof SEQ ID NO: 1, optionally wherein the human FGF21 protein variantcomprises the amino acid sequence of SEQ ID NO: 11. In one embodimentthereof, the human FGF21 variant is provided at a dose of approximately100 mg, 150 mg, 200 mg, 250 mg, or 300 mg for administration once perweek. In one embodiment thereof, the human FGF21 variant is provided ata dose of approximately 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg foradministration once every two weeks. In one embodiment thereof, thehuman FGF21 variant is provided at a dose of approximately 100 mg, 150mg, 200 mg, 250 mg, or 300 mg for administration once every three weeks.In one embodiment thereof, the human FGF21 variant is provided at a doseof approximately 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg foradministration once every four weeks or once per month.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing type 2 diabetes in ahuman subject, wherein the human FGF21 protein variant is provided foradministration at a dose or an amount in the range of 100 mg, 110 mg,120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg,210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg,300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, optionally whereinthe human FGF21 protein variant is provided at a dose of approximately100 mg, 150 mg, 200 mg, 250 mg, or 300 mg, and wherein the human FGF21variant is a fusion protein comprising a human Fc region fused to amature human FGF21 protein or a fragment thereof comprising one or moremutations selected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G,and G202A according to the numbering of SEQ ID NO: 1, optionally whereinthe human FGF21 protein variant comprises the amino acid sequence of SEQID NO: 11. In one embodiment thereof, the human FGF21 variant isprovided at a dose of approximately 100 mg, 150 mg, 200 mg, 250 mg, or300 mg for administration once per week. In one embodiment thereof, thehuman FGF21 variant is provided at a dose of approximately 100 mg, 150mg, 200 mg, 250 mg, or 300 mg for administration once every two weeks.In one embodiment thereof, the human FGF21 variant is provided at a doseof approximately 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg foradministration once every three weeks. In one embodiment thereof, thehuman FGF21 variant is provided at a dose of approximately 100 mg, 150mg, 200 mg, 250 mg, or 300 mg for administration once every four weeksor once per month.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing obesity in a humansubject, wherein the human FGF21 protein variant is provided foradministration at a dose or an amount in the range of 100 mg, 110 mg,120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg,210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg,300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, optionally whereinthe human FGF21 protein variant is provided at a dose of approximately100 mg, 150 mg, 200 mg, 250 mg, or 300 mg, and wherein the human FGF21variant is a fusion protein comprising a human Fc region fused to amature human FGF21 protein or a fragment thereof comprising one or moremutations selected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G,and G202A according to the numbering of SEQ ID NO: 1, optionally whereinthe human FGF21 protein variant comprises the amino acid sequence of SEQID NO: 11. In one embodiment thereof, the human FGF21 variant isprovided at a dose of approximately 100 mg, 150 mg, 200 mg, 250 mg, or300 mg for administration once per week. In one embodiment thereof, thehuman FGF21 variant is provided at a dose of approximately 100 mg, 150mg, 200 mg, 250 mg, or 300 mg for administration once every two weeks.In one embodiment thereof, the human FGF21 variant is provided at a doseof approximately 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg foradministration once every three weeks. In one embodiment thereof, thehuman FGF21 variant is provided at a dose of approximately 100 mg, 150mg, 200 mg, 250 mg, or 300 mg for administration once every four weeksor once per month.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing hypercholesterolemia,dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease(NAFLD), nonalcoholic steatohepatitis (NASH), type 2 diabetes, orobesity in a human subject, wherein the human FGF21 protein variant isprovided for administration at a dose or an amount in the range of 100mg, and wherein the human FGF21 variant is a fusion protein comprising ahuman Fc region fused to a mature human FGF21 protein or a fragmentthereof comprising one or more mutations selected from: Q55C, R105K,G148C, K150R, P158S, S195A, P199G, and G202A according to the numberingof SEQ ID NO: 1, optionally wherein the human FGF21 protein variantcomprises the amino acid sequence of SEQ ID NO: 11. In one embodimentthereof, the human FGF21 variant is provided for administration once perweek. In one embodiment thereof, the human FGF21 variant is provided foradministration once every two weeks. In one embodiment thereof, thehuman FGF21 variant is provided for administration once every threeweeks. In one embodiment thereof, the human FGF21 variant is providedfor administration once every four weeks or once per month.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing hypercholesterolemia,dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease(NAFLD), nonalcoholic steatohepatitis (NASH), type 2 diabetes, orobesity in a human subject, wherein the human FGF21 protein variant isprovided for administration at a dose or an amount in the range of 150mg, and wherein the human FGF21 variant is a fusion protein comprising ahuman Fc region fused to a mature human FGF21 protein or a fragmentthereof comprising one or more mutations selected from: Q55C, R105K,G148C, K150R, P158S, S195A, P199G, and G202A according to the numberingof SEQ ID NO: 1, optionally wherein the human FGF21 protein variantcomprises the amino acid sequence of SEQ ID NO: 11. In one embodimentthereof, the human FGF21 variant is provided for administration once perweek. In one embodiment thereof, the human FGF21 variant is provided foradministration once every two weeks. In one embodiment thereof, thehuman FGF21 variant is provided for administration once every threeweeks. In one embodiment thereof, the human FGF21 variant is providedfor administration once every four weeks or once per month.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing hypercholesterolemia,dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease(NAFLD), nonalcoholic steatohepatitis (NASH), type 2 diabetes, orobesity in a human subject, wherein the human FGF21 protein variant isprovided for administration at a dose or an amount in the range of 200mg, and wherein the human FGF21 variant is a fusion protein comprising ahuman Fc region fused to a mature human FGF21 protein or a fragmentthereof comprising one or more mutations selected from: Q55C, R105K,G148C, K150R, P158S, S195A, P199G, and G202A according to the numberingof SEQ ID NO: 1, optionally wherein the human FGF21 protein variantcomprises the amino acid sequence of SEQ ID NO: 11. In one embodimentthereof, the human FGF21 variant is provided for administration once perweek. In one embodiment thereof, the human FGF21 variant is provided foradministration once every two weeks. In one embodiment thereof, thehuman FGF21 variant is provided for administration once every threeweeks. In one embodiment thereof, the human FGF21 variant is providedfor administration once every four weeks or once per month.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing hypercholesterolemia,dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease(NAFLD), nonalcoholic steatohepatitis (NASH), type 2 diabetes, orobesity in a human subject, wherein the human FGF21 protein variant isprovided for administration at a dose or an amount in the range of 250mg, and wherein the human FGF21 variant is a fusion protein comprising ahuman Fc region fused to a mature human FGF21 protein or a fragmentthereof comprising one or more mutations selected from: Q55C, R105K,G148C, K150R, P158S, S195A, P199G, and G202A according to the numberingof SEQ ID NO: 1, optionally wherein the human FGF21 protein variantcomprises the amino acid sequence of SEQ ID NO: 11. In one embodimentthereof, the human FGF21 variant is provided for administration once perweek. In one embodiment thereof, the human FGF21 variant is provided foradministration once every two weeks. In one embodiment thereof, thehuman FGF21 variant is provided for administration once every threeweeks. In one embodiment thereof, the human FGF21 variant is providedfor administration once every four weeks or once per month.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholicsteatohepatitis (NASH) in a human subject, wherein the human FGF21protein variant is provided for administration at a dose or an amount inthe range of 300 mg, and wherein the human FGF21 variant is a fusionprotein comprising a human Fc region fused to a mature human FGF21protein or a fragment thereof comprising one or more mutations selectedfrom: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202Aaccording to the numbering of SEQ ID NO: 1, optionally wherein the humanFGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the human FGF21 variant is provided foradministration once per week. In one embodiment thereof, the human FGF21variant is provided for administration once every two weeks. In oneembodiment thereof, the human FGF21 variant is provided foradministration once every three weeks. In one embodiment thereof, thehuman FGF21 variant is provided for administration once every four weeksor once per month.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing hypercholesterolemia,dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease(NAFLD), nonalcoholic steatohepatitis (NASH), type 2 diabetes, orobesity in a human subject, wherein the human FGF21 protein variant isprovided for administration at a dose or an amount in the range of 100mg or 150 mg once per week, and wherein the human FGF21 variant is afusion protein comprising a human Fc region fused to a mature humanFGF21 protein or a fragment thereof comprising one or more mutationsselected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202Aaccording to the numbering of SEQ ID NO: 1, optionally wherein the humanFGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing hypercholesterolemia,dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease(NAFLD), nonalcoholic steatohepatitis (NASH), type 2 diabetes, orobesity in a human subject, wherein the human FGF21 protein variant isprovided for administration at a dose or an amount in the range of 150mg or 200 mg once every two weeks, and wherein the human FGF21 variantis a fusion protein comprising a human Fc region fused to a mature humanFGF21 protein or a fragment thereof comprising one or more mutationsselected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202Aaccording to the numbering of SEQ ID NO: 1, optionally wherein the humanFGF21 protein variant comprises the amino acid sequence of SEQ ID NO: 11

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing hypercholesterolemia,dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease(NAFLD), nonalcoholic steatohepatitis (NASH), type 2 diabetes, orobesity in a human subject, wherein the human FGF21 protein variant isprovided for administration at a dose or an amount in the range of 200mg or 250 mg once every three weeks, and wherein the human FGF21 variantis a fusion protein comprising a human Fc region fused to a mature humanFGF21 protein or a fragment thereof comprising one or more mutationsselected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202Aaccording to the numbering of SEQ ID NO: 1, optionally wherein the humanFGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing hypercholesterolemia,dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease(NAFLD), nonalcoholic steatohepatitis (NASH), type 2 diabetes, orobesity in a human subject, wherein the human FGF21 protein variant isprovided for administration at a dose or an amount in the range of 250mg or 300 mg once every four weeks or once per month, and wherein thehuman FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing hypercholesterolemia ina human subject, wherein the human FGF21 protein variant is provided foradministration in combination with one or more additionaltherapeutically active agents, wherein the one or more additionaltherapeutically active agents is a SGLT inhibitor selected from thegroup consisting of dapagliflozin, empagliflozin, canagliflozin,ertugliflozin, sotagliflozin, tofogliflozin, remogliflozin,luseogliflozin, ipragliflozin, atigliflozin, bexagliflozin,henagliflozin, licogliflozin, and a pharmaceutically acceptable salt ofany of these, and wherein the human FGF21 variant is a fusion proteincomprising a human Fc region fused to a mature human FGF21 protein or afragment thereof comprising one or more mutations selected from: Q55C,R105K, G148C, K150R, P158S, S195A, P199G, and G202A according to thenumbering of SEQ ID NO: 1, optionally wherein the human FGF21 proteinvariant comprises the amino acid sequence of SEQ ID NO: 11. In oneembodiment thereof, the SGLT inhibitor is licogliflozin.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing dyslipidemia in a humansubject, wherein the human FGF21 protein variant is provided foradministration in combination with one or more additionaltherapeutically active agents, wherein the one or more additionaltherapeutically active agents is a SGLT inhibitor selected from thegroup consisting of dapagliflozin, empagliflozin, canagliflozin,ertugliflozin, sotagliflozin, tofogliflozin, remogliflozin,luseogliflozin, ipragliflozin, atigliflozin, bexagliflozin,henagliflozin, licogliflozin, and a pharmaceutically acceptable salt ofany of these, and wherein the human FGF21 variant is a fusion proteincomprising a human Fc region fused to a mature human FGF21 protein or afragment thereof comprising one or more mutations selected from: Q55C,R105K, G148C, K150R, P158S, S195A, P199G, and G202A according to thenumbering of SEQ ID NO: 1, optionally wherein the human FGF21 proteinvariant comprises the amino acid sequence of SEQ ID NO: 11. In oneembodiment thereof, the SGLT inhibitor is licogliflozin.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing hypertriglyceridemia ina human subject, wherein the human FGF21 protein variant is provided foradministration in combination with one or more additionaltherapeutically active agents, wherein the one or more additionaltherapeutically active agents is a SGLT inhibitor selected from thegroup consisting of dapagliflozin, empagliflozin, canagliflozin,ertugliflozin, sotagliflozin, tofogliflozin, remogliflozin,luseogliflozin, ipragliflozin, atigliflozin, bexagliflozin,henagliflozin, licogliflozin, and a pharmaceutically acceptable salt ofany of these, and wherein the human FGF21 variant is a fusion proteincomprising a human Fc region fused to a mature human FGF21 protein or afragment thereof comprising one or more mutations selected from: Q55C,R105K, G148C, K150R, P158S, S195A, P199G, and G202A according to thenumbering of SEQ ID NO: 1, optionally wherein the human FGF21 proteinvariant comprises the amino acid sequence of SEQ ID NO: 11. In oneembodiment thereof, the SGLT inhibitor is licogliflozin.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) in a human subject, wherein the human FGF21protein variant is provided for administration in combination with oneor more additional therapeutically active agents, wherein the one ormore additional therapeutically active agents is a SGLT inhibitorselected from the group consisting of dapagliflozin, empagliflozin,canagliflozin, ertugliflozin, sotagliflozin, tofogliflozin,remogliflozin, luseogliflozin, ipragliflozin, atigliflozin,bexagliflozin, henagliflozin, licogliflozin, and a pharmaceuticallyacceptable salt of any of these, and wherein the human FGF21 variant isa fusion protein comprising a human Fc region fused to a mature humanFGF21 protein or a fragment thereof comprising one or more mutationsselected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202Aaccording to the numbering of SEQ ID NO: 1, optionally wherein the humanFGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the SGLT inhibitor is licogliflozin.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholicsteatohepatitis (NASH) in a human subject, wherein the human FGF21protein variant is provided for administration in combination with oneor more additional therapeutically active agents, wherein the one ormore additional therapeutically active agents is a SGLT inhibitorselected from the group consisting of dapagliflozin, empagliflozin,canagliflozin, ertugliflozin, sotagliflozin, tofogliflozin,remogliflozin, luseogliflozin, ipragliflozin, atigliflozin,bexagliflozin, henagliflozin, licogliflozin, and a pharmaceuticallyacceptable salt of any of these, and wherein the human FGF21 variant isa fusion protein comprising a human Fc region fused to a mature humanFGF21 protein or a fragment thereof comprising one or more mutationsselected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202Aaccording to the numbering of SEQ ID NO: 1, optionally wherein the humanFGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the SGLT inhibitor is licogliflozin.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing type 2 diabetes in ahuman subject, wherein the human FGF21 protein variant is provided foradministration in combination with one or more additionaltherapeutically active agents, wherein the one or more additionaltherapeutically active agents is a SGLT inhibitor selected from thegroup consisting of dapagliflozin, empagliflozin, canagliflozin,ertugliflozin, sotagliflozin, tofogliflozin, remogliflozin,luseogliflozin, ipragliflozin, atigliflozin, bexagliflozin,henagliflozin, licogliflozin, and a pharmaceutically acceptable salt ofany of these, and wherein the human FGF21 variant is a fusion proteincomprising a human Fc region fused to a mature human FGF21 protein or afragment thereof comprising one or more mutations selected from: Q55C,R105K, G148C, K150R, P158S, S195A, P199G, and G202A according to thenumbering of SEQ ID NO: 1, optionally wherein the human FGF21 proteinvariant comprises the amino acid sequence of SEQ ID NO: 11. In oneembodiment thereof, the SGLT inhibitor is licogliflozin.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing obesity in a humansubject, wherein the human FGF21 protein variant is provided foradministration in combination with one or more additionaltherapeutically active agents, wherein the one or more additionaltherapeutically active agents is a SGLT inhibitor selected from thegroup consisting of dapagliflozin, empagliflozin, canagliflozin,ertugliflozin, sotagliflozin, tofogliflozin, remogliflozin,luseogliflozin, ipragliflozin, atigliflozin, bexagliflozin,henagliflozin, licogliflozin, and a pharmaceutically acceptable salt ofany of these, and wherein the human FGF21 variant is a fusion proteincomprising a human Fc region fused to a mature human FGF21 protein or afragment thereof comprising one or more mutations selected from: Q55C,R105K, G148C, K150R, P158S, S195A, P199G, and G202A according to thenumbering of SEQ ID NO: 1, optionally wherein the human FGF21 proteinvariant comprises the amino acid sequence of SEQ ID NO: 11. In oneembodiment thereof, the SGLT inhibitor is licogliflozin.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 100 mg once per week and wherein liver fatcontent of the subject is reduced to within a manageable range or normalrange as determined by a clinician or clinical guidelines or by at leastabout 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the humanFGF21 variant is a fusion protein comprising a human Fc region fused toa mature human FGF21 protein or a fragment thereof comprising one ormore mutations selected from: Q55C, R105K, G148C, K150R, P158S, S195A,P199G, and G202A according to the numbering of SEQ ID NO: 1, optionallywherein the human FGF21 protein variant comprises the amino acidsequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 100 mg once every two weeks and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 100 mg once every three weeks and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 100 mg once every four weeks and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 150 mg once per week and wherein liver fatcontent of the subject is reduced to within a manageable range or normalrange as determined by a clinician or clinical guidelines or by at leastabout 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the humanFGF21 variant is a fusion protein comprising a human Fc region fused toa mature human FGF21 protein or a fragment thereof comprising one ormore mutations selected from: Q55C, R105K, G148C, K150R, P158S, S195A,P199G, and G202A according to the numbering of SEQ ID NO: 1, optionallywherein the human FGF21 protein variant comprises the amino acidsequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 150 mg once every two weeks and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 150 mg once every three weeks and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 150 mg once every four weeks and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 200 mg once per week and wherein liver fatcontent of the subject is reduced to within a manageable range or normalrange as determined by a clinician or clinical guidelines or by at leastabout 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the humanFGF21 variant is a fusion protein comprising a human Fc region fused toa mature human FGF21 protein or a fragment thereof comprising one ormore mutations selected from: Q55C, R105K, G148C, K150R, P158S, S195A,P199G, and G202A according to the numbering of SEQ ID NO: 1, optionallywherein the human FGF21 protein variant comprises the amino acidsequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 200 mg once every two weeks and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 200 mg once every three weeks and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 200 mg once every four weeks and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 250 mg once per week and wherein liver fatcontent of the subject is reduced to within a manageable range or normalrange as determined by a clinician or clinical guidelines or by at leastabout 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the humanFGF21 variant is a fusion protein comprising a human Fc region fused toa mature human FGF21 protein or a fragment thereof comprising one ormore mutations selected from: Q55C, R105K, G148C, K150R, P158S, S195A,P199G, and G202A according to the numbering of SEQ ID NO: 1, optionallywherein the human FGF21 protein variant comprises the amino acidsequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 250 mg once every two weeks and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 250 mg once every three weeks and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 250 mg once every four weeks and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 300 mg once per week and wherein liver fatcontent of the subject is reduced to within a manageable range or normalrange as determined by a clinician or clinical guidelines or by at leastabout 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the humanFGF21 variant is a fusion protein comprising a human Fc region fused toa mature human FGF21 protein or a fragment thereof comprising one ormore mutations selected from: Q55C, R105K, G148C, K150R, P158S, S195A,P199G, and G202A according to the numbering of SEQ ID NO: 1, optionallywherein the human FGF21 protein variant comprises the amino acidsequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 300 mg once every two weeks and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 300 mg once every three weeks and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), whereinthe human FGF21 protein variant is provided for administration at a doseor an amount in the range of 300 mg once every four weeks and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholic fattyliver disease (NAFLD) in a human subject, wherein the human FGF21protein variant is provided for administration in combination withlicogliflozin or a pharmaceutically acceptable salt thereof, and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing nonalcoholicsteatohepatitis (NASH) in a human subject, wherein the human FGF21protein variant is provided for administration in combination withlicogliflozin or a pharmaceutically acceptable salt thereof, and whereinliver fat content of the subject is reduced to within a manageable rangeor normal range as determined by a clinician or clinical guidelines orby at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and whereinthe human FGF21 variant is a fusion protein comprising a human Fc regionfused to a mature human FGF21 protein or a fragment thereof comprisingone or more mutations selected from: Q55C, R105K, G148C, K150R, P158S,S195A, P199G, and G202A according to the numbering of SEQ ID NO: 1,optionally wherein the human FGF21 protein variant comprises the aminoacid sequence of SEQ ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing triglyceridemia,wherein the human FGF21 protein variant is provided for administrationat a dose or an amount in the range of 100 mg once per week, once everytwo weeks, once every three weeks, once every four weeks, or oncemonthly and wherein triglyceride levels in the subject are reduced by atleast about 40% or at least about 50%, and wherein the human FGF21variant is a fusion protein comprising a human Fc region fused to amature human FGF21 protein or a fragment thereof comprising one or moremutations selected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G,and G202A according to the numbering of SEQ ID NO: 1, optionally whereinthe human FGF21 protein variant comprises the amino acid sequence of SEQID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing triglyceridemia,wherein the human FGF21 protein variant is provided for administrationat a dose or an amount in the range of 150 mg once per week, once everytwo weeks, once every three weeks, once every four weeks, or oncemonthly and wherein triglyceride levels in the subject are reduced by atleast about 40% or at least about 50%, and wherein the human FGF21variant is a fusion protein comprising a human Fc region fused to amature human FGF21 protein or a fragment thereof comprising one or moremutations selected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G,and G202A according to the numbering of SEQ ID NO: 1, optionally whereinthe human FGF21 protein variant comprises the amino acid sequence of SEQID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing triglyceridemia,wherein the human FGF21 protein variant is provided for administrationat a dose or an amount in the range of 200 mg once per week, once everytwo weeks, once every three weeks, once every four weeks, or oncemonthly and wherein triglyceride levels in the subject are reduced by atleast about 40% or at least about 50%, and wherein the human FGF21variant is a fusion protein comprising a human Fc region fused to amature human FGF21 protein or a fragment thereof comprising one or moremutations selected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G,and G202A according to the numbering of SEQ ID NO: 1, optionally whereinthe human FGF21 protein variant comprises the amino acid sequence of SEQID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing triglyceridemia,wherein the human FGF21 protein variant is provided for administrationat a dose or an amount in the range of 250 mg once per week, once everytwo weeks, once every three weeks, once every four weeks, or oncemonthly and wherein triglyceride levels in the subject are reduced by atleast about 40% or at least about 50%, and wherein the human FGF21variant is a fusion protein comprising a human Fc region fused to amature human FGF21 protein or a fragment thereof comprising one or moremutations selected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G,and G202A according to the numbering of SEQ ID NO: 1, optionally whereinthe human FGF21 protein variant comprises the amino acid sequence of SEQID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for usein a method of treating, preventing, or managing triglyceridemia,wherein the human FGF21 protein variant is provided for administrationat a dose or an amount in the range of 300 mg once per week, once everytwo weeks, once every three weeks, once every four weeks, or oncemonthly and wherein triglyceride levels in the subject are reduced by atleast about 40% or at least about 50%, and wherein the human FGF21variant is a fusion protein comprising a human Fc region fused to amature human FGF21 protein or a fragment thereof comprising one or moremutations selected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G,and G202A according to the numbering of SEQ ID NO: 1, optionally whereinthe human FGF21 protein variant comprises the amino acid sequence of SEQID NO: 11.

Each of the foregoing aspects and embodiments, as well as other elementsdescribed herein, may be combined in any manner without limitation.

1. A human FGF21 protein variant for use in a method of treating,preventing, or managing a metabolic disorder or a cardiovasculardisorder in a human subject, wherein the human FGF21 protein variant isprovided for administration at a dose in the range of 100 mg to 600 mg.2. The human FGF21 protein variant for use according to claim 1, whereinthe metabolic disorder or cardiovascular disorder is selected fromhypercholesterolemia, dyslipidemia, hypertriglyceridemia, nonalcoholicfatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), type 2diabetes, and obesity.
 3. The human FGF21 protein variant for useaccording to claim 1 or 2, wherein treating, preventing, or managing themetabolic disorder or cardiovascular disorder comprises or ischaracterized by reducing one or more of the following: body weight,liver fat content, elevated LDL-C, total-C, triglyceride, and Apo Blevels in the subject.
 4. The human FGF21 protein variant for useaccording to any one of claims 1 to 3, wherein treating, preventing, ormanaging the metabolic disorder or cardiovascular disorder comprises oris characterized by increasing HDL-C levels in the subject.
 5. The humanFGF21 protein variant for use according to claim 3, wherein treating,preventing, or managing the metabolic disorder or cardiovasculardisorder comprises or is characterized by reducing triglyceride levelsin the subject by at least about 40% or at least about 50%.
 6. The humanFGF21 protein variant for use according to claim 1 or 2, whereintreating, preventing, or managing the metabolic disorder orcardiovascular disorder comprises or is characterized by reducingcardiovascular risk in the subject.
 7. The FGF21 protein variant for useaccording to any one of claims 2 to 6, wherein the metabolic disorder orcardiovascular disorder is dyslipidemia, optionally mixed dyslipidemia,hypertriglyceridemia, optionally severe hypertriglyceridemia, orhypercholesterolemia, optionally primary hypercholesterolemia.
 8. TheFGF21 protein variant for use according to any one of claims 2 to 6,wherein the metabolic disorder or cardiovascular disorder isnonalcoholic fatty liver disease (NAFLD) or nonalcoholic steatohepatitis(NASH).
 9. The human FGF21 protein variant for use according to any oneof the preceding claims, wherein the subject is 18 to 55 years of age.10. The human FGF21 protein variant for use according to any one of thepreceding claims, wherein the subject has a body mass index (BMI) withinthe range of 30 to 45 kg/m², inclusive, with ethnic adjustment ≥27.5 fora subject of Asian descent or Asian ancestry.
 11. The human FGF21protein variant for use according to any one of the preceding claims,wherein the subject has triglyceride levels in the range of 150-500mg/dL (1.69-5.65 mmol/L) when measured prior to administration of thehuman FGF21 protein variant.
 12. The human FGF21 protein variant for useaccording to any one of the preceding claims, wherein the human FGF21variant is a fusion protein comprising a human Fc region fused to amature human FGF21 protein or a fragment thereof comprising one or moremutations selected from: Q55C, R105K, G148C, K150R, P158S, S195A, P199G,and G202A according to the numbering of SEQ ID NO:
 1. 13. The humanFGF21 protein variant for use according to claim 12, wherein the humanFGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.
 14. The human FGF21 protein variant for use according to any one ofthe preceding claims, wherein the human FGF21 protein variant isprovided for administration at a dose of at least 100 mg, 150 mg, 200mg, 250 mg, 300 mg, 350 mg, or 400 mg.
 15. The human FGF21 proteinvariant for use according to any one of the preceding claims, whereinthe human FGF21 protein variant is provided for administration at a doseof approximately 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg,270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350mg, optionally wherein the human FGF21 protein variant is provided at adose of approximately 250 mg or 300 mg.
 16. The human FGF21 proteinvariant for use according to any one of the preceding claims, whereinthe human FGF21 protein variant is provided in combination with one ormore additional therapeutically active agents.
 17. The human FGF21protein variant for use according to claim 16, wherein the one or moreadditional therapeutically active agents is selected from the groupconsisting of compounds useful in obesity therapies, diuretics,beta-blockers, alpha-blockers, ACE inhibitors, Angiotensin II ReceptorBlockers (ARBs), direct renin inhibitors, calcium channel blockers,central agonists, peripheral adrenergic blockers, vasodialators,insulin, alpha-glucosidase inhibitors, biguanides, dopamine agonist,DPP-4 inhibitors, glucagon-like peptides, meglitinides, sodium glucosetransporter (SGLT) inhibitors, sulfonylureas, thiazolidinediones,amylinomimetics, statins, fibrates, aspirin, and anticoagulants.
 18. Thehuman FGF21 protein variant for use according to claim 17, wherein thesodium glucose transporter (SGLT) inhibitor is selected fromdapagliflozin, empagliflozin, canagliflozin, ertugliflozin,sotagliflozin, tofogliflozin, remogliflozin, luseogliflozin,ipragliflozin, atigliflozin, bexagliflozin, henagliflozin,licogliflozin, and a pharmaceutically acceptable salt of any of these.19. The human FGF21 protein variant for use according to any one of thepreceding claims, wherein the human FGF21 protein variant is provided ina form for subcutaneous administration.
 20. The human FGF21 proteinvariant for use according to any one of the preceding claims, whereinthe human FGF21 protein variant is provided for administration once amonth or once every 4 weeks, once every 3 weeks, once every 2 weeks, oronce every week.